Novel human transporter proteins and polynucleotides encoding the same

ABSTRACT

Novel human polynucleotide and polypeptide sequences are disclosed that can be used in therapeutic, diagnostic, and pharmacogenomic applications.

[0001] The present application claims the benefit of U.S. ProvisionalApplication No. 60/298,241, which was filed on Jun. 14, 2001, and isherein incorporated by reference in its entirety.

1. INTRODUCTION

[0002] The present invention relates to the discovery, identification,and characterization of novel human polynucleotides encoding proteinsthat share sequence similarity with mammalian transporter proteins. Theinvention encompasses the described polynucleotides, host cellexpression systems, the encoded proteins, fusion proteins, polypeptidesand peptides, antibodies to the encoded proteins and peptides, andgenetically engineered animals that either lack or overexpress thedisclosed polynucleotides, antagonists and agonists of the proteins, andother compounds that modulate the expression or activity of the proteinsencoded by the disclosed polynucleotides, which can be used fordiagnosis, drug screening, clinical trial monitoring, the treatment ofdiseases and disorders, and cosmetic or nutriceutical applications.

2. BACKGROUND OF THE INVENTION

[0003] Transporter proteins are integral membrane proteins that mediateor facilitate the passage of materials across the lipid bilayer. Giventhat the transport of materials across the membrane can play animportant physiological role, transporter proteins are good drugtargets. Additionally, one of the mechanisms of drug resistance involvesdiseased cells using cellular transporter systems to exportchemotherapeutic agents from the cell. Such mechanisms are particularlyrelevant to cells manifesting resistance to a multiplicity of drugs.

3. SUMMARY OF THE INVENTION

[0004] The present invention relates to the discovery, identification,and characterization of nucleotides that encode novel human proteins,and the corresponding amino acid sequences of these proteins. The novelhuman proteins (NHPs) described for the first time herein sharestructural similarity with mammalian ATP-binding cassette (ABC)transporters, organic ion transporters/symporters, and sodium-glucosecotransporters.

[0005] The novel human nucleic acid sequences described herein encodealternative proteins/open reading frames (ORFs) of 1205 and 1207 aminoacids in length (ABC transporter, SEQ ID NOS: 3 and 4, respectively),and 681, 674, 745 and 738 amino acids in length (sodium/glucose-likecotransporter, SEQ ID NOS: 7, 9 11 and 13, respectively).

[0006] The invention also encompasses agonists and antagonists of thedescribed NHPs, including small molecules, large molecules, mutant NHPs,or portions thereof, that compete with native NHPs, peptides, andantibodies, as well as nucleotide sequences that can be used to inhibitthe expression of the described NHPs (e.g., antisense and ribozymemolecules, and open reading frame or regulatory sequence replacementconstructs) or to enhance the expression of the described NHPs (e.g.,expression constructs that place the described polynucleotide under thecontrol of a strong promoter system), and transgenic animals thatexpress a NHP sequence, or “knock-outs” (which can be conditional) thatdo not express a functional NHP. Knock-out mice can be produced inseveral ways, one of which involves the use of mouse embryonic stem cell(“ES cell”) lines that contain gene trap mutations in a murine homologof at least one of the described NHPs. When the unique NHP sequencesdescribed in SEQ ID NOS: 1-13 are “knocked-out” they provide a method ofidentifying phenotypic expression of the particular gene, as well as amethod of assigning function to previously unknown genes. In addition,animals in which the unique NHP sequences described in SEQ ID NOS: 1-13are “knocked-out” provide an unique source in which to elicit antibodiesto homologous and orthologous proteins, which would have been previouslyviewed by the immune system as “self” and therefore would have failed toelicit significant antibody responses. To these ends, gene trappedknockout ES cells have been generated in murine homologs of certain ofthe described NHPs.

[0007] Additionally, the unique NHP sequences described in SEQ ID NOS:1-13 are useful for the identification of protein coding sequences, andmapping an unique gene to a particular chromosome. These sequencesidentify biologically verified exon splice junctions, as opposed tosplice junctions that may have been bioinformatically predicted fromgenomic sequence alone. The sequences of the present invention are alsouseful as additional DNA markers for restriction fragment lengthpolymorphism (RFLP) analysis, and in forensic biology, particularlygiven the presence of nucleotide polymorphisms within the describedsequences.

[0008] Further, the present invention also relates to processes foridentifying compounds that modulate, i.e., act as agonists orantagonists of, NHP expression and/or NHP activity that utilize purifiedpreparations of the described NHPs and/or NHP products, or cellsexpressing the same. Such compounds can be used as therapeutic agentsfor the treatment of any of a wide variety of symptoms associated withbiological disorders or imbalances.

4. DESCRIPTION OF THE SEQUENCE LISTING AND FIGURES

[0009] The Sequence Listing provides the sequences of the described NHPORFs that encode the described NHP amino acid sequences. SEQ ID NO: 5describes a polynucleotide encoding a NHP ORF along with regions offlanking sequence.

5. DETAILED DESCRIPTION OF THE INVENTION

[0010] The NHPs described for the first time herein are novel proteinsthat can be expressed in, inter alia, human cell lines, bone marrow, andosteocarcinoma cells (SEQ ID NOS: 1-5), or lymph node, kidney, fetalliver, liver, testis, thyroid, adrenal gland, small intestine, uterus,bladder, hypothalamus, fetal kidney, and fetal lung cells (SEQ ID NOS:6-13).

[0011] The present invention encompasses the nucleotides presented inthe Sequence Listing, host cells expressing such nucleotides, theexpression products of such nucleotides, and: (a) nucleotides thatencode mammalian homologs of the described polynucleotides, includingthe specifically described NHPs, and the NHP products; (b) nucleotidesthat encode one or more portions of the NHPs that correspond tofunctional domains, and the polypeptide products specified by suchnucleotide sequences, including, but not limited to, the novel regionsof any active domain(s); (c) isolated nucleotides that encode mutantversions, engineered or naturally occurring, of the described NHPs inwhich all or a part of at least one domain is deleted or altered, andthe polypeptide products specified by such nucleotide sequences,including, but not limited to, soluble proteins and peptides in whichall or a portion of the signal (or one or more hydrophobictransmembrane) sequence is deleted; (d) nucleotides that encode chimericfusion proteins containing all or a portion of a coding region of a NHP,or one of its domains (e.g., a receptor or ligand binding domain,accessory protein/self-association domain, etc.) fused to anotherpeptide or polypeptide; or (e) therapeutic or diagnostic derivatives ofthe described polynucleotides, such as oligonucleotides, antisensepolynucleotides, ribozymes, dsRNA, or gene therapy constructs comprisinga sequence first disclosed in the Sequence Listing.

[0012] As discussed above, the present invention includes the human DNAsequences presented in the Sequence Listing (and vectors comprising thesame), and additionally contemplates any nucleotide sequence encoding acontiguous NHP open reading frame (ORF) that hybridizes to a complementof a DNA sequence presented in the Sequence Listing under highlystringent conditions, e.g., hybridization to filter-bound DNA in 0.5 MNaHPO₄, 7% sodium dodecyl sulfate (SDS), 1 mM EDTA at 65° C., andwashing in 0.1×SSC/0.1% SDS at 68° C. (Ausubel et al., eds., 1989,Current Protocols in Molecular Biology, Vol. I, Green PublishingAssociates, Inc., and John Wiley & Sons, Inc., N.Y., at p. 2.10.3) andencodes a functionally equivalent expression product. Additionallycontemplated are any nucleotide sequences that hybridize to thecomplement of a DNA sequence that encodes and expresses an amino acidsequence presented in the Sequence Listing under moderately stringentconditions, e.g., washing in 0.2×SSC/0.1% SDS at 42° C. (Ausubel et al.,1989, supra), yet still encodes a functionally equivalent NHP product.Functional equivalents of a NHP include naturally occurring NHPs presentin other species, and mutant NHPs, whether naturally occurring orengineered (by site directed mutagenesis, gene shuffling, directedevolution as described in, for example, U.S. Pat. No. 5,837,458). Theinvention also includes degenerate nucleic acid variants of thedisclosed NHP polynucleotide sequences.

[0013] Additionally contemplated are polynucleotides encoding NHP ORFs,or their functional equivalents, encoded by polynucleotide sequencesthat are about 99, 95, 90, or about 85 percent similar or identical tocorresponding regions of the nucleotide sequences of the SequenceListing (as measured by BLAST sequence comparison analysis using, forexample, the GCG sequence analysis package, as described herein, usingstandard default settings).

[0014] The invention also includes nucleic acid molecules, preferablyDNA molecules, that hybridize to, and are therefore the complements of,the described NHP nucleotide sequences. Such hybridization conditionsmay be highly stringent or less highly stringent, as described herein.In instances where the nucleic acid molecules are deoxyoligonucleotides(“DNA oligos”), such molecules are generally about 16 to about 100 baseslong, or about 20 to about 80 bases long, or about 34 to about 45 baseslong, or any variation or combination of sizes represented therein thatincorporate a contiguous region of sequence first disclosed in theSequence Listing. Such oligonucleotides can be used in conjunction withthe polymerase chain reaction (PCR) to screen libraries, isolate clones,and prepare cloning and sequencing templates, etc.

[0015] Alternatively, such NHP oligonucleotides can be used ashybridization probes for screening libraries, and assessing geneexpression patterns (particularly using a microarray or high-throughput“chip” format). Additionally, a series of NHP oligonucleotide sequences,or the complements thereof, can be used to represent all or a portion ofthe described NHP sequences. An oligonucleotide or polynucleotidesequence first disclosed in at least a portion of one or more of thesequences of SEQ ID NOS: 1-13 can be used as a hybridization probe inconjunction with a solid support matrix/substrate (resins, beads,membranes, plastics, polymers, metal or metallized substrates,crystalline or polycrystalline substrates, etc.). Of particular note arespatially addressable arrays (i.e., gene chips, microtiter plates, etc.)of oligonucleotides and polynucleotides, or corresponding oligopeptidesand polypeptides, wherein at least one of the biopolymers present on thespatially addressable array comprises an oligonucleotide orpolynucleotide sequence first disclosed in at least one of the sequencesof SEQ ID NOS: 1-13, or an amino acid sequence encoded thereby. Methodsfor attaching biopolymers to, or synthesizing biopolymers on, solidsupport matrices, and conducting binding studies thereon, are disclosedin, inter alia, U.S. Pat. Nos. 5,700,637, 5,556,752, 5,744,305,4,631,211, 5,445,934, 5,252,743, 4,713,326, 5,424,186, and 4,689,405,the disclosures of which are herein incorporated by reference in theirentirety.

[0016] Addressable arrays comprising sequences first disclosed in SEQ IDNOS: 1-13 can be used to identify and characterize the temporal andtissue specific expression of a gene. These addressable arraysincorporate oligonucleotide sequences of sufficient length to confer therequired specificity, yet be within the limitations of the productiontechnology. The length of these probes is usually within a range ofbetween about 8 to about 2000 nucleotides. Preferably the probes consistof 60 nucleotides, and more preferably 25 nucleotides, from thesequences first disclosed in SEQ ID NOS: 1-13.

[0017] For example, a series of NHP oligonucleotide sequences, or thecomplements thereof, can be used in chip format to represent all or aportion of the described sequences. The oligonucleotides, typicallybetween about 16 to about 40 (or any whole number within the statedrange) nucleotides in length, can partially overlap each other, and/orthe sequence may be represented using oligonucleotides that do notoverlap. Accordingly, the described polynucleotide sequences shalltypically comprise at least about two or three distinct oligonucleotidesequences of at least about 8 nucleotides in length that are each firstdisclosed in the described Sequence Listing. Such oligonucleotidesequences can begin at any nucleotide present within a sequence in theSequence Listing, and proceed in either a sense (5′-to-3′) orientationvis-a-vis the described sequence or in an antisense orientation.

[0018] Microarray-based analysis allows the discovery of broad patternsof genetic activity, providing new understanding of gene functions, andgenerating novel and unexpected insight into transcriptional processesand biological mechanisms. The use of addressable arrays comprisingsequences first disclosed in SEQ ID NOS: 1-13 provides detailedinformation about transcriptional changes involved in a specificpathway, potentially leading to the identification of novel components,or gene functions that manifest themselves as novel phenotypes.

[0019] Probes consisting of sequences first disclosed in SEQ ID NOS:1-13 can also be used in the identification, selection, and validationof novel molecular targets for drug discovery. The use of these uniquesequences permits the direct confirmation of drug targets, andrecognition of drug dependent changes in gene expression that aremodulated through pathways distinct from the intended target of thedrug. These unique sequences therefore also have utility in defining andmonitoring both drug action and toxicity.

[0020] As an example of utility, the sequences first disclosed in SEQ IDNOS: 1-13 can be utilized in microarrays, or other assay formats, toscreen collections of genetic material from patients who have aparticular medical condition. These investigations can also be carriedout using the sequences first disclosed in SEQ ID NOS: 1-13 in silico,and by comparing previously collected genetic databases and thedisclosed sequences using computer software known to those in the art.

[0021] Thus the sequences first disclosed in SEQ ID NOS: 1-13 can beused to identify mutations associated with a particular disease, andalso in diagnostic or prognostic assays.

[0022] Although the presently described sequences have been specificallydescribed using nucleotide sequence, it should be appreciated that eachof the sequences can uniquely be described using any of a wide varietyof additional structural attributes, or combinations thereof. Forexample, a given sequence can be described by the net composition of thenucleotides present within a given region of the sequence, inconjunction with the presence of one or more specific oligonucleotidesequence(s) first disclosed in SEQ ID NOS: 1-13. Alternatively, arestriction map specifying the relative positions of restrictionendonuclease digestion sites, or various palindromic or other specificoligonucleotide sequences, can be used to structurally describe a givensequence. Such restriction maps, which are typically generated by widelyavailable computer programs (e.g., the University of Wisconsin GCGsequence analysis package, SEQUENCHER 3.0, Gene Codes Corp., Ann Arbor,Mich., etc.), can optionally be used in conjunction with one or morediscrete nucleotide sequence(s) present in the sequence that can bedescribed by the relative position of the sequence relative to one ormore additional sequence(s) or one or more restriction sites present inthe disclosed sequence.

[0023] For oligonucleotide probes, highly stringent conditions mayrefer, e.g., to washing in 6×SSC/0.05% sodium pyrophosphate at 37° C.(for 14-base oligos), 48° C. (for 17-base oligos), 55° C. (for 20-baseoligos), and 60° C. (for 23-base oligos). These nucleic acid moleculesmay encode or act as NHP antisense molecules, useful, for example, inNHP gene regulation and/or as antisense primers in amplificationreactions of NHP nucleic acid sequences. With respect to NHP generegulation, such techniques can be used to regulate biologicalfunctions. Further, such sequences may be used as part of ribozymeand/or triple helix sequences that are also useful for NHP generegulation.

[0024] Inhibitory antisense or double stranded oligonucleotides canadditionally comprise at least one modified base moiety that is selectedfrom the group including, but not limited to, 5-fluorouracil,5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine,4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil,5-carboxymethylaminomethyl-2-thiouridine,5-carboxymethylaminomethyluracil, dihydrouracil,beta-D-galactosylqueosine, inosine, N6-isopentenyladenine,1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine,2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine,7-methylguanine, 5-methylaminomethyluracil,5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine,5′-methoxycarboxymethyluracil, 5-methoxyuracil,2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v),wybutoxosine, pseudouracil, queosine, 2-thiocytosine,5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil,uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v),5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w,and 2,6-diaminopurine.

[0025] The antisense oligonucleotide can also comprise at least onemodified sugar moiety selected from the group including, but not limitedto, arabinose, 2-fluoroarabinose, xylulose, and hexose.

[0026] In yet another embodiment, the antisense oligonucleotide willcomprise at least one modified phosphate backbone selected from thegroup including, but not limited to, a phosphorothioate, aphosphorodithioate, a phosphoramidothioate, a phosphoramidate, aphosphordiamidate, a methylphosphonate, an alkyl phosphotriester, and aformacetal or analog thereof.

[0027] In yet another embodiment, the antisense oligonucleotide is anα-anomeric oligonucleotide. An α-anomeric oligonucleotide forms specificdouble-stranded hybrids with complementary RNA in which, contrary to theusual β-units, the strands run parallel to each other (Gautier et al.,1987, Nucl. Acids Res. 15:6625-6641). The oligonucleotide is a2′-0-methylribonucleotide (Inoue et al., 1987, Nucl. Acids Res.15:6131-6148), or a chimeric RNA-DNA analogue (Inoue et al., 1987, FEBSLett. 215:327-330). Alternatively, double stranded RNA can be used todisrupt the expression and function of a targeted NHP.

[0028] Oligonucleotides of the invention can be synthesized by standardmethods known in the art, e.g., by use of an automated DNA synthesizer(such as are commercially available from Biosearch, Applied Biosystems,etc.). As examples, phosphorothioate oligonucleotides can be synthesizedby the method of Stein et al. (1988, Nucl. Acids Res. 16:3209), andmethylphosphonate oligonucleotides can be prepared by use of controlledpore glass polymer supports (Sarin et al., 1988, Proc. Natl. Acad. Sci.USA 85:7448-7451), etc.

[0029] Low stringency conditions are well-known to those of skill in theart, and will vary predictably depending on the specific organisms fromwhich the library and the labeled sequences are derived. For guidanceregarding such conditions, see, for example, Sambrook et al., 1989,Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, ColdSpring Harbor, N.Y. (and periodic updates thereof), and Ausubel et al.,1989, supra.

[0030] Alternatively, suitably labeled NHP nucleotide probes can be usedto screen a human genomic library using appropriately stringentconditions or by PCR. The identification and characterization of humangenomic clones is helpful for identifying polymorphisms (including, butnot limited to, nucleotide repeats, microsatellite alleles, singlenucleotide polymorphisms, or coding single nucleotide polymorphisms),determining the genomic structure of a given locus/allele, and designingdiagnostic tests. For example, sequences derived from regions adjacentto the intron/exon boundaries of the human gene can be used to designprimers for use in amplification assays to detect mutations within theexons, introns, splice sites (e.g., splice acceptor and/or donor sites),etc., that can be used in diagnostics and pharmacogenomics.

[0031] For example, the present sequences can be used in restrictionfragment length polymorphism (RFLP) analysis to identify specificindividuals. In this technique, an individual's genomic DNA is digestedwith one or more restriction enzymes, and probed on a Southern blot toyield unique bands for identification (as generally described in U.S.Pat. No. 5,272,057, incorporated herein by reference). In addition, thesequences of the present invention can be used to provide polynucleotidereagents, e.g., PCR primers, targeted to specific loci in the humangenome, which can enhance the reliability of DNA-based forensicidentifications by, for example, providing another “identificationmarker” (i.e., another DNA sequence that is unique to a particularindividual). Actual base sequence information can be used foridentification as an accurate alternative to patterns formed byrestriction enzyme generated fragments.

[0032] Further, a NHP gene homolog can be isolated from nucleic acidfrom an organism of interest by performing PCR using two degenerate or“wobble” oligonucleotide primer pools designed on the basis of aminoacid sequences within the NHP products disclosed herein. The templatefor the reaction may be genomic DNA, or total RNA, mRNA, and/or cDNAobtained by reverse transcription of mRNA prepared from human ornon-human cell lines or tissue known to express, or suspected ofexpressing, an allele of a NHP gene.

[0033] The PCR product can be subcloned and sequenced to ensure that theamplified sequences represent the sequence of the desired NHP gene. ThePCR fragment can then be used to isolate a full length cDNA clone by avariety of methods. For example, the amplified fragment can be labeledand used to screen a cDNA library, such as a bacteriophage cDNA library.Alternatively, the labeled fragment can be used to isolate genomicclones via the screening of a genomic library.

[0034] PCR technology can also be used to isolate full length cDNAsequences. For example, RNA can be isolated, following standardprocedures, from an appropriate cellular or tissue source (i.e., oneknown to express, or suspected of expressing, a NHP gene). A reversetranscription (RT) reaction can be performed on the RNA using anoligonucleotide primer specific for the most 5′ end of the amplifiedfragment for the priming of first strand synthesis. The resultingRNA/DNA hybrid may then be “tailed” using a standard terminaltransferase reaction, the hybrid may be digested with RNase H, andsecond strand synthesis may then be primed with a complementary primer.Thus, cDNA sequences upstream of the amplified fragment can be isolated.For a review of cloning strategies that can be used, see, e.g., Sambrooket al., 1989, supra.

[0035] A cDNA encoding a mutant NHP sequence can be isolated, forexample, by using PCR. In this case, the first cDNA strand may besynthesized by hybridizing an oligo-dT oligonucleotide to mRNA isolatedfrom tissue known to express, or suspected of expressing, a NHP, in anindividual putatively carrying a mutant NHP allele, and by extending thenew strand with reverse transcriptase. The second strand of the cDNA isthen synthesized using an oligonucleotide that hybridizes specificallyto the 5′ end of the normal sequence. Using these two primers, theproduct is then amplified via PCR, optionally cloned into a suitablevector, and subjected to DNA sequence analysis through methodswell-known to those of skill in the art. By comparing the DNA sequenceof the mutant NHP allele to that of a corresponding normal NHP allele,the mutation(s) responsible for the loss or alteration of function ofthe mutant NHP gene product can be ascertained.

[0036] Alternatively, a genomic library can be constructed using DNAobtained from an individual suspected of carrying, or known to carry, amutant NHP allele (e.g., a person manifesting a NHP-associated phenotypesuch as, for example, obesity, high blood pressure, connective tissuedisorders, infertility, etc.), or a cDNA library can be constructedusing RNA from a tissue known to express, or suspected of expressing, amutant NHP allele. A normal NHP gene, or any suitable fragment thereof,can then be labeled and used as a probe to identify the correspondingmutant NHP allele in such libraries. Clones containing mutant NHPsequences can then be purified and subjected to sequence analysisaccording to methods well-known to those skilled in the art.

[0037] Additionally, an expression library can be constructed utilizingcDNA synthesized from, for example, RNA isolated from a tissue known toexpress, or suspected of expressing, a mutant NHP allele in anindividual suspected of carrying, or known to carry, such a mutantallele. In this manner, gene products made by the putatively mutanttissue can be expressed and screened using standard antibody screeningtechniques in conjunction with antibodies raised against a normal NHPproduct, as described below (for screening techniques, see, for example,Harlow and Lane, eds., 1988, “Antibodies: A Laboratory Manual”, ColdSpring Harbor Press, Cold Spring Harbor, N.Y.).

[0038] Additionally, screening can be accomplished by screening withlabeled NHP fusion proteins, such as, for example, alkalinephosphatase-NHP or NHP-alkaline phosphatase fusion proteins. In caseswhere a NHP mutation results in an expression product with alteredfunction (e.g., as a result of a missense or a frameshift mutation),polyclonal antibodies to a NHP are likely to cross-react with acorresponding mutant NHP expression product. Library clones detected viatheir reaction with such labeled antibodies can be purified andsubjected to sequence analysis according to methods well-known in theart.

[0039] The invention also encompasses: (a) DNA vectors that contain anyof the foregoing NHP coding sequences and/or their complements (i.e.,antisense); (b) DNA expression vectors that contain any of the foregoingNHP coding sequences operatively associated with a regulatory elementthat directs the expression of the coding sequences (for example,baculovirus as described in U.S. Pat. No. 5,869,336, herein incorporatedby reference); (c) genetically engineered host cells that contain any ofthe foregoing NHP coding sequences operatively associated with aregulatory element that directs the expression of the coding sequencesin the host cell; and (d) genetically engineered host cells that expressan endogenous NHP sequence under the control of an exogenouslyintroduced regulatory element (i.e., gene activation). As used herein,regulatory elements include, but are not limited to, inducible andnon-inducible promoters, enhancers, operators, and other elements knownto those skilled in the art that drive and regulate expression. Suchregulatory elements include, but are not limited to, the cytomegalovirus(hCMV) immediate early gene, regulatable, viral elements (particularlyretroviral LTR promoters), the early or late promoters of SV40 oradenovirus, the lac system, the trp system, the TAC system, the TRCsystem, the major operator and promoter regions of phage lambda, thecontrol regions of fd coat protein, the promoter for 3-phosphoglyceratekinase (PGK), the promoters of acid phosphatase, and the promoters ofthe yeast α-mating factors.

[0040] The present invention also encompasses antibodies andanti-idiotypic antibodies (including Fab fragments), antagonists andagonists of a NHP, as well as compounds or nucleotide constructs thatinhibit expression of a NHP sequence (transcription factor inhibitors,antisense and ribozyme molecules, or open reading frame sequence orregulatory sequence replacement constructs), or promote the expressionof a NHP (e.g., expression constructs in which NHP coding sequences areoperatively associated with expression control elements such aspromoters, promoter/enhancers, etc.).

[0041] The NHPs or NHP peptides, NHP fusion proteins, NHP nucleotidesequences, antibodies, antagonists and agonists can be useful for thedetection of mutant NHPs, or inappropriately expressed NHPs, for thediagnosis of disease. The NHP proteins or peptides, NHP fusion proteins,NHP nucleotide sequences, host cell expression systems, antibodies,antagonists, agonists and genetically engineered cells and animals canbe used for screening for drugs (or high throughput screening ofcombinatorial libraries) effective in the treatment of the symptomaticor phenotypic manifestations of perturbing the normal function of a NHPin the body. The use of engineered host cells and/or animals may offeran advantage in that such systems allow not only for the identificationof compounds that bind to the endogenous receptor for a NHP, but canalso identify compounds that trigger NHP-mediated activities orpathways.

[0042] Finally, the NHP products can be used as therapeutics. Forexample, soluble derivatives such as NHP peptides/domains correspondingto NHPs, NHP fusion protein products (especially NHP-Ig fusion proteins,i.e., fusions of a NHP, or a domain of a NHP, to an IgFc), NHPantibodies and anti-idiotypic antibodies (including Fab fragments),antagonists or agonists (including compounds that modulate or act ondownstream targets in a NHP-mediated pathway) can be used to directlytreat diseases or disorders. For instance, the administration of aneffective amount of a soluble NHP, a NHP-IgFc fusion protein, or ananti-idiotypic antibody (or its Fab) that mimics the NHP, could activateor effectively antagonize an endogenous NHP receptor. Nucleotideconstructs encoding such NHP products can be used to geneticallyengineer host cells to express such products in vivo; these geneticallyengineered cells function as “bioreactors” in the body delivering acontinuous supply of a NHP, a NHP peptide, or a NHP fusion protein tothe body. Nucleotide constructs encoding functional NHPs, mutant NHPs,as well as antisense and ribozyme molecules can also be used in “genetherapy” approaches for the modulation of NHP expression. Thus, theinvention also encompasses pharmaceutical formulations and methods fortreating biological disorders.

[0043] Various aspects of the invention are described in greater detailin the subsections below.

[0044] 5.1 The NHP Sequences

[0045] The cDNA sequences and the corresponding deduced amino acidsequences of the described NHPs are presented in the Sequence Listing.The NHP nucleotides were obtained from clustered human genomicsequences, and human cDNAs made from bone marrow and trachea mRNA (SEQID NOS: 1-5), while SEQ ID NOS: 6-13 were generated using cDNAsgenerated from human lymph node, thyroid, adrenal gland, uterus, andsmall intestine mRNAs (Edge Biosystems, Gaithersburg, Md., Clontech,Palo Alto, Calif.).

[0046] A number of polymorphisms were identified during the sequencingof the NHPs, including: a T/C polymorphism at the nucleotide positionrepresented by, for example, position 462 of SEQ ID NO: 1 (or position468 of SEQ ID NO: 2), both of which result in a leu at the regioncorresponding to amino acid (aa) position 154 of, for example, SEQ IDNO: 3 (or position 156 of SEQ ID NO: 4); a G/A polymorphism at thenucleotide position represented by, for example, position 123 of SEQ IDNO: 6 (and the corresponding location in SEQ ID NOS: 8. 10 and 12), bothof which result in a val at the region corresponding to aa position 41of, for example, SEQ ID NO: 7 (and the corresponding location in SEQ IDNOS: 9, 11 and 13); a G/A polymorphism at the nucleotide positionrepresented by, for example, position 370 of SEQ ID NO: 6 (and thecorresponding location in SEQ ID NOS: 8. 10 and 12), which can result ina val or ile at the region corresponding to aa position 124 of, forexample, SEQ ID NO: 7 (and the corresponding location in SEQ ID NOS: 9,11 and 13); and a G/A polymorphism at the nucleotide positionrepresented by, for example, position 454 of SEQ ID NO: 6 (and thecorresponding location in SEQ ID NOS: 8. 10 and 12), which can result ina val or met at the region corresponding to aa position 152 of, forexample, SEQ ID NO: 7 (and the corresponding location in SEQ ID NOS: 9,11 and 13). As these polymorphisms are coding single nucleotidepolymorphisms (SNPs), they are particularly useful in forensic analysis.

[0047] SEQ ID NOS: 1-5 describe sequences that are similar to, interalia, mammalian ABC transporter proteins, and are apparently encoded onhuman chromosome 7 (see GenBank Accession Number AC073424). SEQ ID NOS:6-13 describe sequences that are similar to, inter alia, mammaliansodium symporter proteins, and are apparently encoded on either humanchromosome 1 or 4 (see GenBank Accession Numbers AL359959 and AC055887).Accordingly, the described sequences are useful for mapping and/ordefining the corresponding coding regions of the human genome andidentifying exon splice junctions.

[0048] An additional application of the described novel humanpolynucleotide sequences is their use in the molecularmutagenesis/evolution of proteins that are at least partially encoded bythe described novel sequences using, for example, polynucleotideshuffling or related methodologies. Such approaches are described inU.S. Pat. Nos. 5,830,721 and 5,837,458, which are herein incorporated byreference in their entirety.

[0049] NHP gene products can also be expressed in transgenic animals.Animals of any species, including, but not limited to, worms, mice,rats, rabbits, guinea pigs, pigs, micro-pigs, birds, goats, andnon-human primates, e.g., baboons, monkeys, and chimpanzees, may be usedto generate NHP transgenic animals.

[0050] Any technique known in the art may be used to introduce a NHPtransgene into animals to produce the founder lines of transgenicanimals. Such techniques include, but are not limited to, pronuclearmicroinjection (Hoppe and Wagner, 1989, U.S. Pat. No. 4,873,191);retrovirus-mediated gene transfer into germ lines (Van der Putten etal., 1985, Proc. Natl. Acad. Sci. USA 82:6148-6152); gene targeting inembryonic stem cells (Thompson et al., 1989, Cell 56:313-321);electroporation of embryos (Lo, 1983, Mol Cell. Biol. 3:1803-1814); andsperm-mediated gene transfer (Lavitrano et al., 1989, Cell 57:717-723);etc. For a review of such techniques, see Gordon, 1989, TransgenicAnimals, Intl. Rev. Cytol. 115:171-229, which is incorporated byreference herein in its entirety.

[0051] The present invention provides for transgenic animals that carrya NHP transgene in all their cells, as well as animals that carry atransgene in some, but not all their cells, i.e., mosaic animals orsomatic cell transgenic animals. A transgene may be integrated as asingle transgene, or in concatamers, e.g., head-to-head tandems orhead-to-tail tandems. A transgene may also be selectively introducedinto and activated in a particular cell-type by following, for example,the teaching of Lasko et al., 1992, Proc. Natl. Acad. Sci. USA89:6232-6236. The regulatory sequences required for such a cell-typespecific activation will depend upon the particular cell-type ofinterest, and will be apparent to those of skill in the art.

[0052] When it is desired that a NHP transgene be integrated into thechromosomal site of the endogenous NHP gene, gene targeting ispreferred. Briefly, when such a technique is to be utilized, vectorscontaining some nucleotide sequences homologous to the endogenous NHPgene are designed for the purpose of integrating, via homologousrecombination with chromosomal sequences, into and disrupting thefunction of the nucleotide sequence of the endogenous NHP gene (i.e.,“knockout” animals).

[0053] The transgene can also be selectively introduced into aparticular cell-type, thus inactivating the endogenous NHP gene in onlythat cell-type, by following, for example, the teaching of Gu et al.,1994, Science 265:103-106. The regulatory sequences required for such acell-type specific inactivation will depend upon the particularcell-type of interest, and will be apparent to those of skill in theart.

[0054] Once transgenic animals have been generated, the expression ofthe recombinant NHP gene may be assayed utilizing standard techniques.Initial screening may be accomplished by Southern blot analysis or PCRtechniques to analyze animal tissues to assay whether integration of thetransgene has taken place. The level of mRNA expression of the transgenein the tissues of the transgenic animals may also be assessed usingtechniques that include, but are not limited to, Northern blot analysisof tissue samples obtained from the animal, in situ hybridizationanalysis, and RT-PCR. Samples of NHP gene-expressing tissue may also beevaluated immunocytochemically using antibodies specific for the NHPtransgene product.

[0055] The present invention also provides for “knock-in” animals.Knock-in animals are those in which a polynucleotide sequence (i.e., agene or a cDNA) that the animal does not naturally have in its genome isinserted in such a way that it is expressed. Examples include, but arenot limited to, a human gene or cDNA used to replace its murine orthologin the mouse, a murine cDNA used to replace the murine gene in themouse, and a human gene or cDNA or murine cDNA that is tagged with areporter construct used to replace the murine ortholog or gene in themouse. Such replacements can occur at the locus of the murine orthologor gene, or at another specific site. Such knock-in animals are usefulfor the in vivo study, testing and validation of, intra alia, human drugtargets, as well as for compounds that are directed at the same, andtherapeutic proteins.

[0056] 5.2 NHPS and NHP Polypeptides

[0057] NHPS, NHP polypeptides, NHP peptide fragments, mutated,truncated, or deleted forms of the NHPs, and/or NHP fusion proteins canbe prepared for a variety of uses. These uses include, but are notlimited to, the generation of antibodies, as reagents in diagnosticassays, for the identification of other cellular gene products relatedto a NHP, and as reagents in assays for screening for compounds that canbe used as pharmaceutical reagents useful in the therapeutic treatmentof mental, biological, or medical disorders and diseases. Given thesimilarity information and expression data, the described NHPs can betargeted (by drugs, oligos, antibodies, etc.) in order to treat disease,or to therapeutically augment the efficacy of, for example,chemotherapeutic agents used in the treatment of breast or prostatecancer.

[0058] The Sequence Listing discloses the amino acid sequences encodedby the described NHP polynucleotides. The NHPs typically displayinitiator methionines in DNA sequence contexts consistent with atranslation initiation site. SEQ ID NOS: 3 and 4 display signal typesequences similar to those often found on membrane proteins; however,all of the described proteins display multiple transmembrane hydrophobicdomains typical of membrane associated proteins.

[0059] The NHP amino acid sequences of the invention include the aminoacid sequence presented in the Sequence Listing, as well as analoguesand derivatives thereof. Further, corresponding NHP homologues fromother species are encompassed by the invention. In fact, any NHP proteinencoded by the NHP nucleotide sequences described herein are within thescope of the invention, as are any novel polynucleotide sequencesencoding all or any novel portion of an amino acid sequence presented inthe Sequence Listing. The degenerate nature of the genetic code iswell-known, and, accordingly, each amino acid presented in the SequenceListing is generically representative of the well-known nucleic acid“triplet” codon, or in many cases codons, that can encode the aminoacid. As such, as contemplated herein, the amino acid sequencespresented in the Sequence Listing, when taken together with the geneticcode (see, for example, Table 4-1 at page 109 of “Molecular CellBiology”, 1986, J. Darnell et al., eds., Scientific American Books, NewYork, N.Y., herein incorporated by reference), are genericallyrepresentative of all the various permutations and combinations ofnucleic acid sequences that can encode such amino acid sequences.

[0060] The invention also encompasses proteins that are functionallyequivalent to the NHPs encoded by the presently described nucleotidesequences, as judged by any of a number of criteria, including, but notlimited to, the ability to bind and cleave a substrate of a NHP, theability to effect an identical or complementary downstream pathway, or achange in cellular metabolism (e.g., proteolytic activity, ion flux,tyrosine phosphorylation, etc.). Such functionally equivalent NHPproteins include, but are not limited to, additions or substitutions ofamino acid residues within the amino acid sequence encoded by the NHPnucleotide sequences described herein, but that result in a silentchange, thus producing a functionally equivalent expression product.Amino acid substitutions may be made on the basis of similarity inpolarity, charge, solubility, hydrophobicity, hydrophilicity, and/or theamphipathic nature of the residues involved. For example, nonpolar(hydrophobic) amino acids include alanine, leucine, isoleucine, valine,proline, phenylalanine, tryptophan, and methionine; polar neutral aminoacids include glycine, serine, threonine, cysteine, tyrosine,asparagine, and glutamine; positively charged (basic) amino acidsinclude arginine, lysine, and histidine; and negatively charged (acidic)amino acids include aspartic acid and glutamic acid.

[0061] A variety of host-expression vector systems can be used toexpress the NHP nucleotide sequences of the invention. Where, as in thepresent instance, the NHP peptide or polypeptide is thought to be from amembrane protein, the hydrophobic regions of the protein can be excised,and the resulting soluble peptide or polypeptide can be recovered fromthe culture media. Such expression systems also encompass engineeredhost cells that express a NHP, or functional equivalent, in situ.Purification or enrichment of a NHP from such expression systems can beaccomplished using appropriate detergents and lipid micelles and methodswell-known to those skilled in the art. However, such engineered hostcells themselves may be used in situations where it is important notonly to retain the structural and functional characteristics of a NHP,but to assess biological activity, e.g., in certain drug screeningassays.

[0062] The expression systems that may be used for purposes of theinvention include, but are not limited to, microorganisms such asbacteria (e.g., E. coli, B. subtilis) transformed with recombinantbacteriophage DNA, plasmid DNA or cosmid DNA expression vectorscontaining NHP nucleotide sequences; yeast (e.g., Saccharomyces, Pichia)transformed with recombinant yeast expression vectors containing NHPnucleotide sequences; insect cell systems infected with recombinantvirus expression vectors (e.g., baculovirus) containing NHP nucleotidesequences; plant cell systems infected with recombinant virus expressionvectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus,TMV) or transformed with recombinant plasmid expression vectors (e.g.,Ti plasmid) containing NHP nucleotide sequences; or mammalian cellsystems (e.g., COS, CHO, BHK, 293, 3T3) harboring recombinant expressionconstructs containing NHP nucleotide sequences and promoters derivedfrom the genome of mammalian cells (e.g., metallothionein promoter) orfrom mammalian viruses (e.g., the adenovirus late promoter; the vacciniavirus 7.5K promoter).

[0063] In bacterial systems, a number of expression vectors may beadvantageously selected depending upon the use intended for the NHPproduct being expressed. For example, when a large quantity of such aprotein is to be produced for the generation of pharmaceuticalcompositions of or containing a NHP, or for raising antibodies to a NHP,vectors that direct the expression of high levels of fusion proteinproducts that are readily purified may be desirable. Such vectorsinclude, but are not limited to, the E. coli expression vector pUR278(Ruther et al., 1983, EMBO J. 2:1791), in which a NHP coding sequencemay be ligated individually into the vector in-frame with the lacZcoding region so that a fusion protein is produced; pIN vectors (Inouyeand Inouye, 1985, Nucleic Acids Res. 13:3101-3109; Van Heeke andSchuster, 1989, J. Biol. Chem. 264:5503-5509); and the like. pGEXvectors (Pharmacia or American Type Culture Collection) can also be usedto express foreign polypeptides as fusion proteins with glutathioneS-transferase (GST). In general, such fusion proteins are soluble andcan easily be purified from lysed cells by adsorption toglutathione-agarose beads, followed by elution in the presence of freeglutathione. The pGEX vectors are designed to include thrombin or factorXa protease cleavage sites so that the cloned target expression productcan be released from the GST moiety.

[0064] In an exemplary insect system, Autographa californica nuclearpolyhedrosis virus (AcNPV) is used as a vector to express foreignpolynucleotide sequences. The virus grows in Spodoptera frugiperdacells. A NHP coding sequence can be cloned individually into anon-essential region (for example the polyhedrin gene) of the virus andplaced under control of an AcNPV promoter (for example the polyhedrinpromoter). Successful insertion of a NHP coding sequence will result ininactivation of the polyhedrin gene and production of non-occludedrecombinant virus (i.e., virus lacking the proteinaceous coat coded forby the polyhedrin gene). These recombinant viruses are then used toinfect Spodoptera frugiperda cells in which the inserted sequence isexpressed (e.g., see Smith et al., 1983, J. Virol. 46:584; Smith, U.S.Pat. No. 4,215,051).

[0065] In mammalian host cells, a number of viral-based expressionsystems may be utilized. In cases where an adenovirus is used as anexpression vector, the NHP nucleotide sequence of interest may beligated to an adenovirus transcription/translation control complex,e.g., the late promoter and tripartite leader sequence. This chimericsequence may then be inserted in the adenovirus genome by in vitro or invivo recombination. Insertion in a non-essential region of the viralgenome (e.g., region E1 or E3) will result in a recombinant virus thatis viable and capable of expressing a NHP product in infected hosts(e.g., see Logan and Shenk, 1984, Proc. Natl. Acad. Sci. USA81:3655-3659). Specific initiation signals may also be required forefficient translation of inserted NHP nucleotide sequences. Thesesignals include the ATG initiation codon and adjacent sequences. Incases where an entire NHP gene or cDNA, including its own initiationcodon and adjacent sequences, is inserted into the appropriateexpression vector, no additional translational control signals may beneeded. However, in cases where only a portion of a NHP coding sequenceis inserted, exogenous translational control signals, including,perhaps, the ATG initiation codon, may be provided. Furthermore, theinitiation codon should be in phase with the reading frame of thedesired coding sequence to ensure translation of the entire insert.These exogenous translational control signals and initiation codons canbe of a variety of origins, both natural and synthetic. The efficiencyof expression may be enhanced by the inclusion of appropriatetranscription enhancer elements, transcription terminators, etc. (seeBitter et al., 1987, Methods in Enzymol. 153:516-544).

[0066] In addition, a host cell strain may be chosen that modulates theexpression of the inserted sequences, or modifies and processes theexpression product in the specific fashion desired. Such modifications(e.g., glycosylation) and processing (e.g., cleavage) of proteinproducts may be important for the function of the protein. Differenthost cells have characteristic and specific mechanisms for thepost-translational processing and modification of proteins andexpression products. Appropriate cell lines or host systems can bechosen to ensure the desired modification and processing of the foreignprotein expressed. To this end, eukaryotic host cells that possess thecellular machinery for the desired processing of the primary transcript,glycosylation, and phosphorylation of the expression product may beused. Such mammalian host cells include, but are not limited to, CHO,VERO, BHK, HeLa, COS, MDCK, 293, 3T3, WI38, and in particular, humancell lines.

[0067] For long-term, high-yield production of recombinant proteins,stable expression is preferred. For example, cell lines that stablyexpress the NHP sequences described herein can be engineered. Ratherthan using expression vectors that contain viral origins of replication,host cells can be transformed with DNA controlled by appropriateexpression control elements (e.g., promoter, enhancer sequences,transcription terminators, polyadenylation sites, etc.), and aselectable marker. Following the introduction of the foreign DNA,engineered cells may be allowed to grow for 1-2 days in an enrichedmedia, and then switched to a selective media. The selectable marker inthe recombinant plasmid confers resistance to the selection and allowscells to stably integrate the plasmid into their chromosomes and grow toform foci, which in turn can be cloned and expanded into cell lines.This method may advantageously be used to engineer cell lines thatexpress a NHP product. Such engineered cell lines may be particularlyuseful in screening and evaluation of compounds that affect theendogenous activity of a NHP product.

[0068] A number of selection systems may be used, including, but notlimited to, the herpes simplex virus thymidine kinase (Wigler et al.,1977, Cell 11:223), hypoxanthine-guanine phosphoribosyltransferase(Szybalska and Szybalski, 1962, Proc. Natl. Acad. Sci. USA 48:2026), andadenine phosphoribosyltransferase (Lowy et al., 1980, Cell 22:817)genes, which can be employed in tk⁻, hgprt⁻ or aprt⁻ cells,respectively. Also, antimetabolite resistance can be used as the basisof selection for the following genes: dhfr, which confers resistance tomethotrexate (Wigler et al., 1980, Proc. Natl. Acad. Sci. USA 77:3567;O'Hare et al., 1981, Proc. Natl. Acad. Sci. USA 78:1527); gpt, whichconfers resistance to mycophenolic acid (Mulligan and Berg, 1981, Proc.Natl. Acad. Sci. USA 78:2072); neo, which confers resistance to theaminoglycoside G-418 (Colberre-Garapin et al., 1981, J. Mol. Biol.150:1); and hygro, which confers resistance to hygromycin (Santerre etal., 1984, Gene 30:147).

[0069] Alternatively, any fusion protein can be readily purified byutilizing an antibody specific for the fusion protein being expressed.Another exemplary system allows for the ready purification ofnon-denatured fusion proteins expressed in human cell lines (Janknechtet al., 1991, Proc. Natl. Acad. Sci. USA 88:8972-8976). In this system,the sequence of interest is subcloned into a vaccinia recombinationplasmid such that the sequence's open reading frame is translationallyfused to an amino-terminal tag consisting of six histidine is residues.Extracts from cells infected with recombinant vaccinia virus are loadedonto Ni²⁺.nitriloacetic acid-agarose columns, and histidine-taggedproteins are selectively eluted with imidazole-containing buffers.

[0070] Also encompassed by the present invention are fusion proteinsthat direct a NHP to a target organ and/or facilitate transport acrossthe membrane into the cytosol. Conjugation of NHPs to antibody moleculesor their Fab fragments could be used to target cells bearing aparticular epitope. Attaching an appropriate signal sequence to a NHPwould also transport a NHP to a desired location within the cell.Alternatively targeting of a NHP or its nucleic acid sequence might beachieved using liposome or lipid complex based delivery systems. Suchtechnologies are described in “Liposomes: A Practical Approach”, New,R.R.C., ed., Oxford University Press, N.Y., and in U.S. Pat. Nos.4,594,595, 5,459,127, 5,948,767 and 6,110,490 and their respectivedisclosures, which are herein incorporated by reference in theirentirety. Additionally embodied are novel protein constructs engineeredin such a way that they facilitate transport of NHPs to a target site ordesired organ, where they cross the cell membrane and/or the nucleuswhere the NHPs can exert their functional activity. This goal may beachieved by coupling of a NHP to a cytokine or other ligand thatprovides targeting specificity, and/or to a protein transducing domain(see generally U.S. Provisional Patent Application Ser. Nos. 60/111,701and 60/056,713, both of which are herein incorporated by reference, forexamples of such transducing sequences), to facilitate passage acrosscellular membranes, and can optionally be engineered to include nuclearlocalization signals.

[0071] Additionally contemplated are oligopeptides that are modeled onan amino acid sequence first described in the Sequence Listing. Such NHPoligopeptides are generally between about 10 to about 100 amino acidslong, or between about 16 to about 80 amino acids long, or between about20 to about 35 amino acids long, or any variation or combination ofsizes represented therein that incorporate a contiguous region ofsequence first disclosed in the Sequence Listing. Such NHP oligopeptidescan be of any length disclosed within the above ranges, and can initiateat any amino acid position represented in the Sequence Listing.

[0072] The invention also contemplates “substantially isolated” or“substantially pure” proteins or polypeptides. By a “substantiallyisolated” or “substantially pure” protein or polypeptide is meant aprotein or polypeptide that has been separated from at least some ofthose components that naturally accompany it. Typically, the protein orpolypeptide is substantially isolated or pure when it is at least 60%,by weight, free from the proteins and other naturally-occurring organicmolecules with which it is naturally associated in vivo. Preferably, thepurity of the preparation is at least 75%, more preferably at least 90%,and most preferably at least 99%, by weight. A substantially isolated orpure protein or polypeptide may be obtained, for example, by extractionfrom a natural source, by expression of a recombinant nucleic acidencoding the protein or polypeptide, or by chemically synthesizing theprotein or polypeptide.

[0073] Purity can be measured by any appropriate method, e.g., columnchromatography such as immunoaffinity chromatography using an antibodyspecific for the protein or polypeptide, polyacrylamide gelelectrophoresis, or HPLC analysis. A protein or polypeptide issubstantially free of naturally associated components when it isseparated from at least some of those contaminants that accompany it inits natural state. Thus, a polypeptide that is chemically synthesized orproduced in a cellular system different from the cell from which itnaturally originates will be, by definition, substantially free from itsnaturally associated components. Accordingly, substantially isolated orpure proteins or polypeptides include eukaryotic proteins synthesized inE. coli, other prokaryotes, or any other organism in which they do notnaturally occur.

[0074] 5.3 Antibodies to NHP Products

[0075] Antibodies that specifically recognize one or more epitopes of aNHP, epitopes of conserved variants of a NHP, or peptide fragments of aNHP, are also encompassed by the invention. Such antibodies include, butare not limited to, polyclonal antibodies, monoclonal antibodies (mAbs),humanized or chimeric antibodies, single chain antibodies, Fabfragments, F(ab′)₂ fragments, fragments produced by a Fab expressionlibrary, anti-idiotypic (anti-Id) antibodies, and epitope-bindingfragments of any of the above.

[0076] The antibodies of the invention may be used, for example, in thedetection of a NHP in a biological sample and may, therefore, beutilized as part of a diagnostic or prognostic technique wherebypatients may be tested for abnormal amounts of a NHP. Such antibodiesmay also be utilized in conjunction with, for example, compoundscreening schemes for the evaluation of the effect of test compounds onexpression and/or activity of a NHP expression product. Additionally,such antibodies can be used in conjunction with gene therapy to, forexample, evaluate normal and/or engineered NHP-expressing cells prior totheir introduction into a patient. Such antibodies may additionally beused in methods for the inhibition of abnormal NHP activity. Thus, suchantibodies may be utilized as a part of treatment methods.

[0077] For the production of antibodies, various host animals may beimmunized by injection with a NHP, a NHP peptide (e.g., onecorresponding to a functional domain of a NHP), a truncated NHPpolypeptide (a NHP in which one or more domains have been deleted),functional equivalents of a NHP or mutated variants of a NHP. Such hostanimals may include, but are not limited to, pigs, rabbits, mice, goats,and rats, to name but a few. Various adjuvants may be used to increasethe immunological response, depending on the host species, including,but not limited to, Freund's adjuvant (complete and incomplete), mineralsalts such as aluminum hydroxide or aluminum phosphate, chitosan,surface active substances such as lysolecithin, pluronic polyols,polyanions, peptides, oil emulsions, and potentially useful humanadjuvants such as BCG (bacille Calmette-Guerin) and Corynebacteriumparvum. Alternatively, the immune response could be enhanced bycombination and/or coupling with molecules such as keyhole limpethemocyanin, tetanus toxoid, diphtheria toxoid, ovalbumin, cholera toxin,or fragments thereof. Polyclonal antibodies are heterogeneouspopulations of antibody molecules derived from the sera of the immunizedanimals.

[0078] Monoclonal antibodies, which are homogeneous populations ofantibodies to a particular antigen, can be obtained by any techniquethat provides for the production of antibody molecules by continuouscell lines in culture. These include, but are not limited to, thehybridoma technique of Kohler and Milstein, (1975, Nature 256:495-497;and U.S. Pat. No. 4,376,110), the human B-cell hybridoma technique(Kosbor et al., 1983, Immunology Today 4:72; Cole et al., 1983, Proc.Natl. Acad. Sci. USA 80:2026-2030), and the EBV-hybridoma technique(Cole et al., 1985, Monoclonal Antibodies And Cancer Therapy, Alan R.Liss, Inc., pp. 77-96). Such antibodies may be of any immunoglobulinclass, including IgG, IgM, IgE, IgA, and IgD, and any subclass thereof.The hybridomas producing the mAbs of this invention may be cultivated invitro or in vivo. Production of high titers of mAbs in vivo makes thisthe presently preferred method of production.

[0079] In addition, techniques developed for the production of “chimericantibodies” (Morrison et al., 1984, Proc. Natl. Acad. Sci. USA81:6851-6855; Neuberger et al., 1984, Nature, 312:604-608; Takeda etal., 1985, Nature, 314:452-454) by splicing the genes from a mouseantibody molecule of appropriate antigen specificity together with genesfrom a human antibody molecule of appropriate biological activity can beused. A chimeric antibody is a molecule in which different portions arederived from different animal species, such as those having a variableregion derived from a murine mAb and a human immunoglobulin constantregion. Such technologies are described in U.S. Pat. Nos. 6,114,598,6,075,181 and 5,877,397 and their respective disclosures, which areherein incorporated by reference in their entirety. Also encompassed bythe present invention is the use of fully humanized monoclonalantibodies, as described in U.S. Pat. No. 6,150,584 and respectivedisclosures, which are herein incorporated by reference in theirentirety.

[0080] Alternatively, techniques described for the production of singlechain antibodies (U.S. Pat. No. 4,946,778; Bird, 1988, Science242:423-426; Huston et al., 1988, Proc. Natl. Acad. Sci. USA85:5879-5883; and Ward et al., 1989, Nature 341:544-546) can be adaptedto produce single chain antibodies against NHP expression products.Single chain antibodies are formed by linking the heavy and light chainfragments of the Fv region via an amino acid bridge, resulting in asingle chain polypeptide.

[0081] Antibody fragments that recognize specific epitopes may begenerated by known techniques. For example, such fragments include, butare not limited to: F(ab′)₂ fragments, which can be produced by pepsindigestion of an antibody molecule; and Fab fragments, which can begenerated by reducing the disulfide bridges of F(ab′)₂ fragments.Alternatively, Fab expression libraries may be constructed (Huse et al.,1989, Science, 246:1275-1281) to allow rapid and easy identification ofmonoclonal Fab fragments with the desired specificity.

[0082] Antibodies to a NHP can, in turn, be utilized to generateanti-idiotype antibodies that “mimic” a given NHP, using techniqueswell-known to those skilled in the art (see, e.g., Greenspan and Bona,1993, FASEB J. 7:437-444; and Nissinoff, 1991, J. Immunol.147:2429-2438). For example, antibodies that bind to a NHP domain andcompetitively inhibit the binding of a NHP to its cognate receptor canbe used to generate anti-idiotypes that “mimic” the NHP and, therefore,bind and activate or neutralize a receptor. Such anti-idiotypicantibodies, or Fab fragments of such anti-idiotypes, can be used intherapeutic regimens involving a NHP-mediated pathway.

[0083] Additionally, given the high degree of relatedness of mammalianNHPs, the presently described knock-out mice (having never seen a NHP,and thus never been tolerized to a NHP) have an unique utility, as theycan be advantageously applied to the generation of antibodies againstthe disclosed mammalian NHPs (i.e., a NHP will be immunogenic in NHPknock-out animals).

[0084] The present invention is not to be limited in scope by thespecific embodiments described herein, which are intended as singleillustrations of individual aspects of the invention, and functionallyequivalent methods and components are within the scope of the invention.Indeed, various modifications of the invention, in addition to thoseshown and described herein, will become apparent to those skilled in theart from the foregoing description. Such modifications are intended tofall within the scope of the appended claims. All cited publications,patents, and patent applications are herein incorporated by reference intheir entirety.

1 13 1 3618 DNA homo sapiens 1 atgggttgca catttttacc cttttatgtcattgtatata tttttttgct aagtgttgtt 60 gagatttgtg aagttttcca gcagactgtgaagccctcag aagccatgga gatgctgcag 120 aaagtgaaga tgatggtcgt acgtgtgctcaccatcgttg cagaaaaccc ttcctggacc 180 aaggacattt tgtgtgctac tctgagttgcaagcaaaatg ggataaggca tctcatttta 240 tctgctatac aaggggtcac tttggcgcaggaccacttcc aggaaattga aaagatatgg 300 tcctcgccga atcagctaaa ttgtgaaagtcttagcaaga atctttctag caccttggag 360 agcttcaaga gcagcttgga aaatgccactggccaggact gcacaagcca gccgaggctg 420 gagacggtgc agcagcactt gtacatgttggccaaaagcc tygaggaaac ttggtcatca 480 gggaatccca tcatgacttt tctcagcaatttcacagtaa ctgaggatgt aaaaataaaa 540 gatttgatga agaatatcac caagttgactgaggagcttc gctcttccat ccaaatctcg 600 aatgagacta tccatagcat tctagaagcaaatatttccc actccaaggt tctcttcagt 660 gccctcaccg tagctctgtc tggaaagtgtgatcaggaaa tccttcatct cctgctgaca 720 tttcccaaag gggaaaaatc ttggatcgcagcggaggaac tctgtagcct gccagggtca 780 aaagtgtatt ctctgattgt gttgctgagtcgaaacttgg atgtgcgagc tttcatttac 840 aagactctga tgccttctga agcaaatggcttgctcaact ccttgctgga tatagtttcc 900 agcctcagcg ccttgcttgc caaagcccagcacgtctttg agtatcttcc tgagtttctt 960 cacacattta aaatcactgc cttgctagaaaccctggact ttcaacaggt ttcacaaaat 1020 gtccaggcca gaagttcagc ttttggttctttccagtttg tgatgaagat ggtttgcaag 1080 gaccaagcat cattccttag cgattctaatatgtttatta atttgcccag agttaaggaa 1140 ctcttggaag atgacaaaga aaaattcaacattcctgaag attcaacacc gttttgcttg 1200 aagctttatc aggaaattct acaattgccaaatggtgctt tggtgtggac cttcctaaaa 1260 cccatattgc atggaaaaat actatacacaccaaacactc cagaaattaa caaggtcatt 1320 caaaaggcta attacacctt ttatattgtggacaaactaa aaactttatc agaaacactg 1380 ctggaaatgt ccagcctttt ccagagaagtggaagtggcc agatgttcaa ccagctgcag 1440 gaggccctga gaaacaaatt tgtaagaaactttgtagaaa accagttgca cattgatgta 1500 gacaaactta ctgaaaaact ccagacatacggagggctgc tggatgagat gtttaaccat 1560 gcaggcgctg gacgcttccg tttcttgggcagcatcttgg tcaatctctc ttcctgcgtg 1620 gcactgaacc gtttccaggc tctgcagtctgtcgacatcc tggagactaa agcacatgaa 1680 ctcttgcagc agaacagctt cttggccagtatcattttca gcaattcctt attcgacaag 1740 aacttcagat cagagtctgt caaactgccaccccatgtct catacacaat ccggaccaat 1800 gtgttataca gcgtgcgaac agatgtggtaaaaaaccctt cttggaagtt ccaccctcag 1860 aatctaccag ctgatgggtt caaatataactacgtctttg ccccactgca agacatgatc 1920 gaaagagcca tcattttggt gcagactgggcaggaagccc tggaaccagc agcacagact 1980 caggcggccc cttacccctg ccataccagcgacctattcc tgaacaacgt tggtttcttt 2040 tttccactga taatgatgct gacgtggatggtgtctgtgg ccagcatggt cagaaagttg 2100 gtgtatgagc aggagataca gatagaagagtatatgcgga tgatgggagt gcatccagtg 2160 atccatttcc tggcctggtt cctggagaacatggctgtgt tgaccataag cagtgctact 2220 ctggccatcg ttctgaaaac aagtggcatctttgcacaca gcaatacctt tattgttttc 2280 ctctttctct tggattttgg gatgtcagtcgtcatgctga gctacctctt gagtgcattt 2340 ttcagccaag ctaatacagc ggccctttgtaccagcctgg tgtacatgat cagctttctg 2400 ccctacatag ttctattggt tctacataaccaattaagtt ttgttaatca gacatttctg 2460 tgccttcttt cgacaaccgc ctttggacaaggggtatttt ttattacatt cctggaagga 2520 caagagacag ggattcaatg gaataatatgtaccaggctc tggaacaagg gggcatgaca 2580 tttggctggg tttgctggat gattctttttgattcaagcc tttatttttt gtgtggatgg 2640 tacttgagca acttgattcc tggaacatttggtttacgga aaccatggta tttccccttt 2700 actgcctcat attggaagag tgtgggtttcttggtggaga aaaggcaata ctttctaagt 2760 tctagtctgt tcttcttcaa tgagaactttgacaataaag ggtcatcact gcaaaacagg 2820 gaaggagagc ttgaaggaag tgccccgggagtcaccctgg tgtctgtgac caaggaatat 2880 gagggccaca aggctgtggt ccaagacctcagcctgacct tctacagaga ccaaatcacc 2940 gccctgctgg ggacaaacgg tgccgggaaaaccactatca tatccatgtt gacggggctc 3000 caccctccca cttctggaac catcatcatcaatggcaaga acctacagac agacctgtcg 3060 agggtcagaa tggagcttgg tgtgtgtccgcagcaggaca tcctgttgga caacctcacc 3120 gtccgggaac atttgctgct ctttgcttccataaaggcgc ctcagtggac caagaaggag 3180 ctgcatcagc aagtcaatca aactcttcaggatgtggact taactcagca tcagcacaaa 3240 cagacccgag ctctgtctgg aggcctgaagaggaagctct cccttggcat tgctttcatg 3300 ggcatgtcga ggaccgtggt tctggatgagcccaccagtg gggtggaccc ttgctcccgg 3360 catagcctgt gggacattct gctcaagtaccgagaaggta ggcactgggc ctcattctgc 3420 cttctcttcc cacaatattg tgttgcaggaaatgcattgc tactgtacag tagaatcaag 3480 ttgtatccca gtgaggctac attatccttttcagaaaaat ataaattttt aaaagcactt 3540 atagggatat attcgttaga taacatctctatagtgctta gaattgctta ctttgtgttt 3600 gaccttttaa ctcaataa 3618 2 3624DNA homo sapiens 2 atgcacatgg gttgcacatt tttacccttt tatgtcattgtatatatttt tttgctaagt 60 gttgttgaga tttgtgaagt tttccagcag actgtgaagccctcagaagc catggagatg 120 ctgcagaaag tgaagatgat ggtcgtacgt gtgctcaccatcgttgcaga aaacccttcc 180 tggaccaagg acattttgtg tgctactctg agttgcaagcaaaatgggat aaggcatctc 240 attttatctg ctatacaagg ggtcactttg gcgcaggaccacttccagga aattgaaaag 300 atatggtcct cgccgaatca gctaaattgt gaaagtcttagcaagaatct ttctagcacc 360 ttggagagct tcaagagcag cttggaaaat gccactggccaggactgcac aagccagccg 420 aggctggaga cggtgcagca gcacttgtac atgttggccaaaagcctyga ggaaacttgg 480 tcatcaggga atcccatcat gacttttctc agcaatttcacagtaactga ggatgtaaaa 540 ataaaagatt tgatgaagaa tatcaccaag ttgactgaggagcttcgctc ttccatccaa 600 atctcgaatg agactatcca tagcattcta gaagcaaatatttcccactc caaggttctc 660 ttcagtgccc tcaccgtagc tctgtctgga aagtgtgatcaggaaatcct tcatctcctg 720 ctgacatttc ccaaagggga aaaatcttgg atcgcagcggaggaactctg tagcctgcca 780 gggtcaaaag tgtattctct gattgtgttg ctgagtcgaaacttggatgt gcgagctttc 840 atttacaaga ctctgatgcc ttctgaagca aatggcttgctcaactcctt gctggatata 900 gtttccagcc tcagcgcctt gcttgccaaa gcccagcacgtctttgagta tcttcctgag 960 tttcttcaca catttaaaat cactgccttg ctagaaaccctggactttca acaggtttca 1020 caaaatgtcc aggccagaag ttcagctttt ggttctttccagtttgtgat gaagatggtt 1080 tgcaaggacc aagcatcatt ccttagcgat tctaatatgtttattaattt gcccagagtt 1140 aaggaactct tggaagatga caaagaaaaa ttcaacattcctgaagattc aacaccgttt 1200 tgcttgaagc tttatcagga aattctacaa ttgccaaatggtgctttggt gtggaccttc 1260 ctaaaaccca tattgcatgg aaaaatacta tacacaccaaacactccaga aattaacaag 1320 gtcattcaaa aggctaatta caccttttat attgtggacaaactaaaaac tttatcagaa 1380 acactgctgg aaatgtccag ccttttccag agaagtggaagtggccagat gttcaaccag 1440 ctgcaggagg ccctgagaaa caaatttgta agaaactttgtagaaaacca gttgcacatt 1500 gatgtagaca aacttactga aaaactccag acatacggagggctgctgga tgagatgttt 1560 aaccatgcag gcgctggacg cttccgtttc ttgggcagcatcttggtcaa tctctcttcc 1620 tgcgtggcac tgaaccgttt ccaggctctg cagtctgtcgacatcctgga gactaaagca 1680 catgaactct tgcagcagaa cagcttcttg gccagtatcattttcagcaa ttccttattc 1740 gacaagaact tcagatcaga gtctgtcaaa ctgccaccccatgtctcata cacaatccgg 1800 accaatgtgt tatacagcgt gcgaacagat gtggtaaaaaacccttcttg gaagttccac 1860 cctcagaatc taccagctga tgggttcaaa tataactacgtctttgcccc actgcaagac 1920 atgatcgaaa gagccatcat tttggtgcag actgggcaggaagccctgga accagcagca 1980 cagactcagg cggcccctta cccctgccat accagcgacctattcctgaa caacgttggt 2040 ttcttttttc cactgataat gatgctgacg tggatggtgtctgtggccag catggtcaga 2100 aagttggtgt atgagcagga gatacagata gaagagtatatgcggatgat gggagtgcat 2160 ccagtgatcc atttcctggc ctggttcctg gagaacatggctgtgttgac cataagcagt 2220 gctactctgg ccatcgttct gaaaacaagt ggcatctttgcacacagcaa tacctttatt 2280 gttttcctct ttctcttgga ttttgggatg tcagtcgtcatgctgagcta cctcttgagt 2340 gcatttttca gccaagctaa tacagcggcc ctttgtaccagcctggtgta catgatcagc 2400 tttctgccct acatagttct attggttcta cataaccaattaagttttgt taatcagaca 2460 tttctgtgcc ttctttcgac aaccgccttt ggacaaggggtattttttat tacattcctg 2520 gaaggacaag agacagggat tcaatggaat aatatgtaccaggctctgga acaagggggc 2580 atgacatttg gctgggtttg ctggatgatt ctttttgattcaagccttta ttttttgtgt 2640 ggatggtact tgagcaactt gattcctgga acatttggtttacggaaacc atggtatttc 2700 ccctttactg cctcatattg gaagagtgtg ggtttcttggtggagaaaag gcaatacttt 2760 ctaagttcta gtctgttctt cttcaatgag aactttgacaataaagggtc atcactgcaa 2820 aacagggaag gagagcttga aggaagtgcc ccgggagtcaccctggtgtc tgtgaccaag 2880 gaatatgagg gccacaaggc tgtggtccaa gacctcagcctgaccttcta cagagaccaa 2940 atcaccgccc tgctggggac aaacggtgcc gggaaaaccactatcatatc catgttgacg 3000 gggctccacc ctcccacttc tggaaccatc atcatcaatggcaagaacct acagacagac 3060 ctgtcgaggg tcagaatgga gcttggtgtg tgtccgcagcaggacatcct gttggacaac 3120 ctcaccgtcc gggaacattt gctgctcttt gcttccataaaggcgcctca gtggaccaag 3180 aaggagctgc atcagcaagt caatcaaact cttcaggatgtggacttaac tcagcatcag 3240 cacaaacaga cccgagctct gtctggaggc ctgaagaggaagctctccct tggcattgct 3300 ttcatgggca tgtcgaggac cgtggttctg gatgagcccaccagtggggt ggacccttgc 3360 tcccggcata gcctgtggga cattctgctc aagtaccgagaaggtaggca ctgggcctca 3420 ttctgccttc tcttcccaca atattgtgtt gcaggaaatgcattgctact gtacagtaga 3480 atcaagttgt atcccagtga ggctacatta tccttttcagaaaaatataa atttttaaaa 3540 gcacttatag ggatatattc gttagataac atctctatagtgcttagaat tgcttacttt 3600 gtgtttgacc ttttaactca ataa 3624 3 1205 PRThomo sapiens 3 Met Gly Cys Thr Phe Leu Pro Phe Tyr Val Ile Val Tyr IlePhe Leu 1 5 10 15 Leu Ser Val Val Glu Ile Cys Glu Val Phe Gln Gln ThrVal Lys Pro 20 25 30 Ser Glu Ala Met Glu Met Leu Gln Lys Val Lys Met MetVal Val Arg 35 40 45 Val Leu Thr Ile Val Ala Glu Asn Pro Ser Trp Thr LysAsp Ile Leu 50 55 60 Cys Ala Thr Leu Ser Cys Lys Gln Asn Gly Ile Arg HisLeu Ile Leu 65 70 75 80 Ser Ala Ile Gln Gly Val Thr Leu Ala Gln Asp HisPhe Gln Glu Ile 85 90 95 Glu Lys Ile Trp Ser Ser Pro Asn Gln Leu Asn CysGlu Ser Leu Ser 100 105 110 Lys Asn Leu Ser Ser Thr Leu Glu Ser Phe LysSer Ser Leu Glu Asn 115 120 125 Ala Thr Gly Gln Asp Cys Thr Ser Gln ProArg Leu Glu Thr Val Gln 130 135 140 Gln His Leu Tyr Met Leu Ala Lys SerLeu Glu Glu Thr Trp Ser Ser 145 150 155 160 Gly Asn Pro Ile Met Thr PheLeu Ser Asn Phe Thr Val Thr Glu Asp 165 170 175 Val Lys Ile Lys Asp LeuMet Lys Asn Ile Thr Lys Leu Thr Glu Glu 180 185 190 Leu Arg Ser Ser IleGln Ile Ser Asn Glu Thr Ile His Ser Ile Leu 195 200 205 Glu Ala Asn IleSer His Ser Lys Val Leu Phe Ser Ala Leu Thr Val 210 215 220 Ala Leu SerGly Lys Cys Asp Gln Glu Ile Leu His Leu Leu Leu Thr 225 230 235 240 PhePro Lys Gly Glu Lys Ser Trp Ile Ala Ala Glu Glu Leu Cys Ser 245 250 255Leu Pro Gly Ser Lys Val Tyr Ser Leu Ile Val Leu Leu Ser Arg Asn 260 265270 Leu Asp Val Arg Ala Phe Ile Tyr Lys Thr Leu Met Pro Ser Glu Ala 275280 285 Asn Gly Leu Leu Asn Ser Leu Leu Asp Ile Val Ser Ser Leu Ser Ala290 295 300 Leu Leu Ala Lys Ala Gln His Val Phe Glu Tyr Leu Pro Glu PheLeu 305 310 315 320 His Thr Phe Lys Ile Thr Ala Leu Leu Glu Thr Leu AspPhe Gln Gln 325 330 335 Val Ser Gln Asn Val Gln Ala Arg Ser Ser Ala PheGly Ser Phe Gln 340 345 350 Phe Val Met Lys Met Val Cys Lys Asp Gln AlaSer Phe Leu Ser Asp 355 360 365 Ser Asn Met Phe Ile Asn Leu Pro Arg ValLys Glu Leu Leu Glu Asp 370 375 380 Asp Lys Glu Lys Phe Asn Ile Pro GluAsp Ser Thr Pro Phe Cys Leu 385 390 395 400 Lys Leu Tyr Gln Glu Ile LeuGln Leu Pro Asn Gly Ala Leu Val Trp 405 410 415 Thr Phe Leu Lys Pro IleLeu His Gly Lys Ile Leu Tyr Thr Pro Asn 420 425 430 Thr Pro Glu Ile AsnLys Val Ile Gln Lys Ala Asn Tyr Thr Phe Tyr 435 440 445 Ile Val Asp LysLeu Lys Thr Leu Ser Glu Thr Leu Leu Glu Met Ser 450 455 460 Ser Leu PheGln Arg Ser Gly Ser Gly Gln Met Phe Asn Gln Leu Gln 465 470 475 480 GluAla Leu Arg Asn Lys Phe Val Arg Asn Phe Val Glu Asn Gln Leu 485 490 495His Ile Asp Val Asp Lys Leu Thr Glu Lys Leu Gln Thr Tyr Gly Gly 500 505510 Leu Leu Asp Glu Met Phe Asn His Ala Gly Ala Gly Arg Phe Arg Phe 515520 525 Leu Gly Ser Ile Leu Val Asn Leu Ser Ser Cys Val Ala Leu Asn Arg530 535 540 Phe Gln Ala Leu Gln Ser Val Asp Ile Leu Glu Thr Lys Ala HisGlu 545 550 555 560 Leu Leu Gln Gln Asn Ser Phe Leu Ala Ser Ile Ile PheSer Asn Ser 565 570 575 Leu Phe Asp Lys Asn Phe Arg Ser Glu Ser Val LysLeu Pro Pro His 580 585 590 Val Ser Tyr Thr Ile Arg Thr Asn Val Leu TyrSer Val Arg Thr Asp 595 600 605 Val Val Lys Asn Pro Ser Trp Lys Phe HisPro Gln Asn Leu Pro Ala 610 615 620 Asp Gly Phe Lys Tyr Asn Tyr Val PheAla Pro Leu Gln Asp Met Ile 625 630 635 640 Glu Arg Ala Ile Ile Leu ValGln Thr Gly Gln Glu Ala Leu Glu Pro 645 650 655 Ala Ala Gln Thr Gln AlaAla Pro Tyr Pro Cys His Thr Ser Asp Leu 660 665 670 Phe Leu Asn Asn ValGly Phe Phe Phe Pro Leu Ile Met Met Leu Thr 675 680 685 Trp Met Val SerVal Ala Ser Met Val Arg Lys Leu Val Tyr Glu Gln 690 695 700 Glu Ile GlnIle Glu Glu Tyr Met Arg Met Met Gly Val His Pro Val 705 710 715 720 IleHis Phe Leu Ala Trp Phe Leu Glu Asn Met Ala Val Leu Thr Ile 725 730 735Ser Ser Ala Thr Leu Ala Ile Val Leu Lys Thr Ser Gly Ile Phe Ala 740 745750 His Ser Asn Thr Phe Ile Val Phe Leu Phe Leu Leu Asp Phe Gly Met 755760 765 Ser Val Val Met Leu Ser Tyr Leu Leu Ser Ala Phe Phe Ser Gln Ala770 775 780 Asn Thr Ala Ala Leu Cys Thr Ser Leu Val Tyr Met Ile Ser PheLeu 785 790 795 800 Pro Tyr Ile Val Leu Leu Val Leu His Asn Gln Leu SerPhe Val Asn 805 810 815 Gln Thr Phe Leu Cys Leu Leu Ser Thr Thr Ala PheGly Gln Gly Val 820 825 830 Phe Phe Ile Thr Phe Leu Glu Gly Gln Glu ThrGly Ile Gln Trp Asn 835 840 845 Asn Met Tyr Gln Ala Leu Glu Gln Gly GlyMet Thr Phe Gly Trp Val 850 855 860 Cys Trp Met Ile Leu Phe Asp Ser SerLeu Tyr Phe Leu Cys Gly Trp 865 870 875 880 Tyr Leu Ser Asn Leu Ile ProGly Thr Phe Gly Leu Arg Lys Pro Trp 885 890 895 Tyr Phe Pro Phe Thr AlaSer Tyr Trp Lys Ser Val Gly Phe Leu Val 900 905 910 Glu Lys Arg Gln TyrPhe Leu Ser Ser Ser Leu Phe Phe Phe Asn Glu 915 920 925 Asn Phe Asp AsnLys Gly Ser Ser Leu Gln Asn Arg Glu Gly Glu Leu 930 935 940 Glu Gly SerAla Pro Gly Val Thr Leu Val Ser Val Thr Lys Glu Tyr 945 950 955 960 GluGly His Lys Ala Val Val Gln Asp Leu Ser Leu Thr Phe Tyr Arg 965 970 975Asp Gln Ile Thr Ala Leu Leu Gly Thr Asn Gly Ala Gly Lys Thr Thr 980 985990 Ile Ile Ser Met Leu Thr Gly Leu His Pro Pro Thr Ser Gly Thr Ile 9951000 1005 Ile Ile Asn Gly Lys Asn Leu Gln Thr Asp Leu Ser Arg Val ArgMet 1010 1015 1020 Glu Leu Gly Val Cys Pro Gln Gln Asp Ile Leu Leu AspAsn Leu Thr 1025 1030 1035 1040 Val Arg Glu His Leu Leu Leu Phe Ala SerIle Lys Ala Pro Gln Trp 1045 1050 1055 Thr Lys Lys Glu Leu His Gln GlnVal Asn Gln Thr Leu Gln Asp Val 1060 1065 1070 Asp Leu Thr Gln His GlnHis Lys Gln Thr Arg Ala Leu Ser Gly Gly 1075 1080 1085 Leu Lys Arg LysLeu Ser Leu Gly Ile Ala Phe Met Gly Met Ser Arg 1090 1095 1100 Thr ValVal Leu Asp Glu Pro Thr Ser Gly Val Asp Pro Cys Ser Arg 1105 1110 11151120 His Ser Leu Trp Asp Ile Leu Leu Lys Tyr Arg Glu Gly Arg His Trp1125 1130 1135 Ala Ser Phe Cys Leu Leu Phe Pro Gln Tyr Cys Val Ala GlyAsn Ala 1140 1145 1150 Leu Leu Leu Tyr Ser Arg Ile Lys Leu Tyr Pro SerGlu Ala Thr Leu 1155 1160 1165 Ser Phe Ser Glu Lys Tyr Lys Phe Leu LysAla Leu Ile Gly Ile Tyr 1170 1175 1180 Ser Leu Asp Asn Ile Ser Ile ValLeu Arg Ile Ala Tyr Phe Val Phe 1185 1190 1195 1200 Asp Leu Leu Thr Gln1205 4 1207 PRT homo sapiens 4 Met His Met Gly Cys Thr Phe Leu Pro PheTyr Val Ile Val Tyr Ile 1 5 10 15 Phe Leu Leu Ser Val Val Glu Ile CysGlu Val Phe Gln Gln Thr Val 20 25 30 Lys Pro Ser Glu Ala Met Glu Met LeuGln Lys Val Lys Met Met Val 35 40 45 Val Arg Val Leu Thr Ile Val Ala GluAsn Pro Ser Trp Thr Lys Asp 50 55 60 Ile Leu Cys Ala Thr Leu Ser Cys LysGln Asn Gly Ile Arg His Leu 65 70 75 80 Ile Leu Ser Ala Ile Gln Gly ValThr Leu Ala Gln Asp His Phe Gln 85 90 95 Glu Ile Glu Lys Ile Trp Ser SerPro Asn Gln Leu Asn Cys Glu Ser 100 105 110 Leu Ser Lys Asn Leu Ser SerThr Leu Glu Ser Phe Lys Ser Ser Leu 115 120 125 Glu Asn Ala Thr Gly GlnAsp Cys Thr Ser Gln Pro Arg Leu Glu Thr 130 135 140 Val Gln Gln His LeuTyr Met Leu Ala Lys Ser Leu Glu Glu Thr Trp 145 150 155 160 Ser Ser GlyAsn Pro Ile Met Thr Phe Leu Ser Asn Phe Thr Val Thr 165 170 175 Glu AspVal Lys Ile Lys Asp Leu Met Lys Asn Ile Thr Lys Leu Thr 180 185 190 GluGlu Leu Arg Ser Ser Ile Gln Ile Ser Asn Glu Thr Ile His Ser 195 200 205Ile Leu Glu Ala Asn Ile Ser His Ser Lys Val Leu Phe Ser Ala Leu 210 215220 Thr Val Ala Leu Ser Gly Lys Cys Asp Gln Glu Ile Leu His Leu Leu 225230 235 240 Leu Thr Phe Pro Lys Gly Glu Lys Ser Trp Ile Ala Ala Glu GluLeu 245 250 255 Cys Ser Leu Pro Gly Ser Lys Val Tyr Ser Leu Ile Val LeuLeu Ser 260 265 270 Arg Asn Leu Asp Val Arg Ala Phe Ile Tyr Lys Thr LeuMet Pro Ser 275 280 285 Glu Ala Asn Gly Leu Leu Asn Ser Leu Leu Asp IleVal Ser Ser Leu 290 295 300 Ser Ala Leu Leu Ala Lys Ala Gln His Val PheGlu Tyr Leu Pro Glu 305 310 315 320 Phe Leu His Thr Phe Lys Ile Thr AlaLeu Leu Glu Thr Leu Asp Phe 325 330 335 Gln Gln Val Ser Gln Asn Val GlnAla Arg Ser Ser Ala Phe Gly Ser 340 345 350 Phe Gln Phe Val Met Lys MetVal Cys Lys Asp Gln Ala Ser Phe Leu 355 360 365 Ser Asp Ser Asn Met PheIle Asn Leu Pro Arg Val Lys Glu Leu Leu 370 375 380 Glu Asp Asp Lys GluLys Phe Asn Ile Pro Glu Asp Ser Thr Pro Phe 385 390 395 400 Cys Leu LysLeu Tyr Gln Glu Ile Leu Gln Leu Pro Asn Gly Ala Leu 405 410 415 Val TrpThr Phe Leu Lys Pro Ile Leu His Gly Lys Ile Leu Tyr Thr 420 425 430 ProAsn Thr Pro Glu Ile Asn Lys Val Ile Gln Lys Ala Asn Tyr Thr 435 440 445Phe Tyr Ile Val Asp Lys Leu Lys Thr Leu Ser Glu Thr Leu Leu Glu 450 455460 Met Ser Ser Leu Phe Gln Arg Ser Gly Ser Gly Gln Met Phe Asn Gln 465470 475 480 Leu Gln Glu Ala Leu Arg Asn Lys Phe Val Arg Asn Phe Val GluAsn 485 490 495 Gln Leu His Ile Asp Val Asp Lys Leu Thr Glu Lys Leu GlnThr Tyr 500 505 510 Gly Gly Leu Leu Asp Glu Met Phe Asn His Ala Gly AlaGly Arg Phe 515 520 525 Arg Phe Leu Gly Ser Ile Leu Val Asn Leu Ser SerCys Val Ala Leu 530 535 540 Asn Arg Phe Gln Ala Leu Gln Ser Val Asp IleLeu Glu Thr Lys Ala 545 550 555 560 His Glu Leu Leu Gln Gln Asn Ser PheLeu Ala Ser Ile Ile Phe Ser 565 570 575 Asn Ser Leu Phe Asp Lys Asn PheArg Ser Glu Ser Val Lys Leu Pro 580 585 590 Pro His Val Ser Tyr Thr IleArg Thr Asn Val Leu Tyr Ser Val Arg 595 600 605 Thr Asp Val Val Lys AsnPro Ser Trp Lys Phe His Pro Gln Asn Leu 610 615 620 Pro Ala Asp Gly PheLys Tyr Asn Tyr Val Phe Ala Pro Leu Gln Asp 625 630 635 640 Met Ile GluArg Ala Ile Ile Leu Val Gln Thr Gly Gln Glu Ala Leu 645 650 655 Glu ProAla Ala Gln Thr Gln Ala Ala Pro Tyr Pro Cys His Thr Ser 660 665 670 AspLeu Phe Leu Asn Asn Val Gly Phe Phe Phe Pro Leu Ile Met Met 675 680 685Leu Thr Trp Met Val Ser Val Ala Ser Met Val Arg Lys Leu Val Tyr 690 695700 Glu Gln Glu Ile Gln Ile Glu Glu Tyr Met Arg Met Met Gly Val His 705710 715 720 Pro Val Ile His Phe Leu Ala Trp Phe Leu Glu Asn Met Ala ValLeu 725 730 735 Thr Ile Ser Ser Ala Thr Leu Ala Ile Val Leu Lys Thr SerGly Ile 740 745 750 Phe Ala His Ser Asn Thr Phe Ile Val Phe Leu Phe LeuLeu Asp Phe 755 760 765 Gly Met Ser Val Val Met Leu Ser Tyr Leu Leu SerAla Phe Phe Ser 770 775 780 Gln Ala Asn Thr Ala Ala Leu Cys Thr Ser LeuVal Tyr Met Ile Ser 785 790 795 800 Phe Leu Pro Tyr Ile Val Leu Leu ValLeu His Asn Gln Leu Ser Phe 805 810 815 Val Asn Gln Thr Phe Leu Cys LeuLeu Ser Thr Thr Ala Phe Gly Gln 820 825 830 Gly Val Phe Phe Ile Thr PheLeu Glu Gly Gln Glu Thr Gly Ile Gln 835 840 845 Trp Asn Asn Met Tyr GlnAla Leu Glu Gln Gly Gly Met Thr Phe Gly 850 855 860 Trp Val Cys Trp MetIle Leu Phe Asp Ser Ser Leu Tyr Phe Leu Cys 865 870 875 880 Gly Trp TyrLeu Ser Asn Leu Ile Pro Gly Thr Phe Gly Leu Arg Lys 885 890 895 Pro TrpTyr Phe Pro Phe Thr Ala Ser Tyr Trp Lys Ser Val Gly Phe 900 905 910 LeuVal Glu Lys Arg Gln Tyr Phe Leu Ser Ser Ser Leu Phe Phe Phe 915 920 925Asn Glu Asn Phe Asp Asn Lys Gly Ser Ser Leu Gln Asn Arg Glu Gly 930 935940 Glu Leu Glu Gly Ser Ala Pro Gly Val Thr Leu Val Ser Val Thr Lys 945950 955 960 Glu Tyr Glu Gly His Lys Ala Val Val Gln Asp Leu Ser Leu ThrPhe 965 970 975 Tyr Arg Asp Gln Ile Thr Ala Leu Leu Gly Thr Asn Gly AlaGly Lys 980 985 990 Thr Thr Ile Ile Ser Met Leu Thr Gly Leu His Pro ProThr Ser Gly 995 1000 1005 Thr Ile Ile Ile Asn Gly Lys Asn Leu Gln ThrAsp Leu Ser Arg Val 1010 1015 1020 Arg Met Glu Leu Gly Val Cys Pro GlnGln Asp Ile Leu Leu Asp Asn 1025 1030 1035 1040 Leu Thr Val Arg Glu HisLeu Leu Leu Phe Ala Ser Ile Lys Ala Pro 1045 1050 1055 Gln Trp Thr LysLys Glu Leu His Gln Gln Val Asn Gln Thr Leu Gln 1060 1065 1070 Asp ValAsp Leu Thr Gln His Gln His Lys Gln Thr Arg Ala Leu Ser 1075 1080 1085Gly Gly Leu Lys Arg Lys Leu Ser Leu Gly Ile Ala Phe Met Gly Met 10901095 1100 Ser Arg Thr Val Val Leu Asp Glu Pro Thr Ser Gly Val Asp ProCys 1105 1110 1115 1120 Ser Arg His Ser Leu Trp Asp Ile Leu Leu Lys TyrArg Glu Gly Arg 1125 1130 1135 His Trp Ala Ser Phe Cys Leu Leu Phe ProGln Tyr Cys Val Ala Gly 1140 1145 1150 Asn Ala Leu Leu Leu Tyr Ser ArgIle Lys Leu Tyr Pro Ser Glu Ala 1155 1160 1165 Thr Leu Ser Phe Ser GluLys Tyr Lys Phe Leu Lys Ala Leu Ile Gly 1170 1175 1180 Ile Tyr Ser LeuAsp Asn Ile Ser Ile Val Leu Arg Ile Ala Tyr Phe 1185 1190 1195 1200 ValPhe Asp Leu Leu Thr Gln 1205 5 4165 DNA homo sapiens 5 tggagacctaaagttttcta aaggtccaga atgtgttatc tgtgttttct tatgttccta 60 tgaagaaaatatgattatgc agggagggag gatggttctt acatgtgtgt tataacttaa 120 cctacacagtagagatgcac atgggttgca catttttacc cttttatgtc attgtatata 180 tttttttgctaagtgttgtt gagatttgtg aagttttcca gcagactgtg aagccctcag 240 aagccatggagatgctgcag aaagtgaaga tgatggtcgt acgtgtgctc accatcgttg 300 cagaaaacccttcctggacc aaggacattt tgtgtgctac tctgagttgc aagcaaaatg 360 ggataaggcatctcatttta tctgctatac aaggggtcac tttggcgcag gaccacttcc 420 aggaaattgaaaagatatgg tcctcgccga atcagctaaa ttgtgaaagt cttagcaaga 480 atctttctagcaccttggag agcttcaaga gcagcttgga aaatgccact ggccaggact 540 gcacaagccagccgaggctg gagacggtgc agcagcactt gtacatgttg gccaaaagcc 600 tygaggaaacttggtcatca gggaatccca tcatgacttt tctcagcaat ttcacagtaa 660 ctgaggatgtaaaaataaaa gatttgatga agaatatcac caagttgact gaggagcttc 720 gctcttccatccaaatctcg aatgagacta tccatagcat tctagaagca aatatttccc 780 actccaaggttctcttcagt gccctcaccg tagctctgtc tggaaagtgt gatcaggaaa 840 tccttcatctcctgctgaca tttcccaaag gggaaaaatc ttggatcgca gcggaggaac 900 tctgtagcctgccagggtca aaagtgtatt ctctgattgt gttgctgagt cgaaacttgg 960 atgtgcgagctttcatttac aagactctga tgccttctga agcaaatggc ttgctcaact 1020 ccttgctggatatagtttcc agcctcagcg ccttgcttgc caaagcccag cacgtctttg 1080 agtatcttcctgagtttctt cacacattta aaatcactgc cttgctagaa accctggact 1140 ttcaacaggtttcacaaaat gtccaggcca gaagttcagc ttttggttct ttccagtttg 1200 tgatgaagatggtttgcaag gaccaagcat cattccttag cgattctaat atgtttatta 1260 atttgcccagagttaaggaa ctcttggaag atgacaaaga aaaattcaac attcctgaag 1320 attcaacaccgttttgcttg aagctttatc aggaaattct acaattgcca aatggtgctt 1380 tggtgtggaccttcctaaaa cccatattgc atggaaaaat actatacaca ccaaacactc 1440 cagaaattaacaaggtcatt caaaaggcta attacacctt ttatattgtg gacaaactaa 1500 aaactttatcagaaacactg ctggaaatgt ccagcctttt ccagagaagt ggaagtggcc 1560 agatgttcaaccagctgcag gaggccctga gaaacaaatt tgtaagaaac tttgtagaaa 1620 accagttgcacattgatgta gacaaactta ctgaaaaact ccagacatac ggagggctgc 1680 tggatgagatgtttaaccat gcaggcgctg gacgcttccg tttcttgggc agcatcttgg 1740 tcaatctctcttcctgcgtg gcactgaacc gtttccaggc tctgcagtct gtcgacatcc 1800 tggagactaaagcacatgaa ctcttgcagc agaacagctt cttggccagt atcattttca 1860 gcaattccttattcgacaag aacttcagat cagagtctgt caaactgcca ccccatgtct 1920 catacacaatccggaccaat gtgttataca gcgtgcgaac agatgtggta aaaaaccctt 1980 cttggaagttccaccctcag aatctaccag ctgatgggtt caaatataac tacgtctttg 2040 ccccactgcaagacatgatc gaaagagcca tcattttggt gcagactggg caggaagccc 2100 tggaaccagcagcacagact caggcggccc cttacccctg ccataccagc gacctattcc 2160 tgaacaacgttggtttcttt tttccactga taatgatgct gacgtggatg gtgtctgtgg 2220 ccagcatggtcagaaagttg gtgtatgagc aggagataca gatagaagag tatatgcgga 2280 tgatgggagtgcatccagtg atccatttcc tggcctggtt cctggagaac atggctgtgt 2340 tgaccataagcagtgctact ctggccatcg ttctgaaaac aagtggcatc tttgcacaca 2400 gcaatacctttattgttttc ctctttctct tggattttgg gatgtcagtc gtcatgctga 2460 gctacctcttgagtgcattt ttcagccaag ctaatacagc ggccctttgt accagcctgg 2520 tgtacatgatcagctttctg ccctacatag ttctattggt tctacataac caattaagtt 2580 ttgttaatcagacatttctg tgccttcttt cgacaaccgc ctttggacaa ggggtatttt 2640 ttattacattcctggaagga caagagacag ggattcaatg gaataatatg taccaggctc 2700 tggaacaagggggcatgaca tttggctggg tttgctggat gattcttttt gattcaagcc 2760 tttattttttgtgtggatgg tacttgagca acttgattcc tggaacattt ggtttacgga 2820 aaccatggtatttccccttt actgcctcat attggaagag tgtgggtttc ttggtggaga 2880 aaaggcaatactttctaagt tctagtctgt tcttcttcaa tgagaacttt gacaataaag 2940 ggtcatcactgcaaaacagg gaaggagagc ttgaaggaag tgccccggga gtcaccctgg 3000 tgtctgtgaccaaggaatat gagggccaca aggctgtggt ccaagacctc agcctgacct 3060 tctacagagaccaaatcacc gccctgctgg ggacaaacgg tgccgggaaa accactatca 3120 tatccatgttgacggggctc caccctccca cttctggaac catcatcatc aatggcaaga 3180 acctacagacagacctgtcg agggtcagaa tggagcttgg tgtgtgtccg cagcaggaca 3240 tcctgttggacaacctcacc gtccgggaac atttgctgct ctttgcttcc ataaaggcgc 3300 ctcagtggaccaagaaggag ctgcatcagc aagtcaatca aactcttcag gatgtggact 3360 taactcagcatcagcacaaa cagacccgag ctctgtctgg aggcctgaag aggaagctct 3420 cccttggcattgctttcatg ggcatgtcga ggaccgtggt tctggatgag cccaccagtg 3480 gggtggacccttgctcccgg catagcctgt gggacattct gctcaagtac cgagaaggta 3540 ggcactgggcctcattctgc cttctcttcc cacaatattg tgttgcagga aatgcattgc 3600 tactgtacagtagaatcaag ttgtatccca gtgaggctac attatccttt tcagaaaaat 3660 ataaatttttaaaagcactt atagggatat attcgttaga taacatctct atagtgctta 3720 gaattgcttactttgtgttt gaccttttaa ctcaataaca gcaatgacat ctatgtacat 3780 tatacattatcatacatgat ttcaaggaaa attgtcttct tctggaagca tagtttctta 3840 gaagaggcatcccagatcat aggacaagcc tcccttgtct cagatgaaga aatgaaggct 3900 cagagagacgggcatgtgat ttacttgtag ctacagagaa agtttcctga actgagggtg 3960 gatgttgaacctcttgtcca tgtttctcac atctattatt gtttctttcc aatttaggac 4020 atttgatgggcagttactaa tttccaactt ctgattcttt ctgcaatcct gacagctagg 4080 aagcattgttctatgtattt tctgtgagaa tactcccttt tggaaagaaa cattgcaaca 4140 gtaaaacacatcttggtgct ggtaa 4165 6 2046 DNA homo sapiens 6 atgagcaagg agctggcagcaatggggcct ggagcttcag gggacggggt caggactgag 60 acagctccac acatagcactggactccaga gttggtctgc acgcctacga catcagcgtg 120 gtrgtcatct actttgtcttcgtcattgct gtggggatct ggtcgtccat ccgtgcaagt 180 cgagggacca ttggcggctatttcctggcc gggaggtcca tgagctggtg gccaattgga 240 gcatctctga tgtccagcaatgtgggcagt ggcttgttca tcggcctggc tgggacaggg 300 gctgccggag gccttgccgtaggtggcttc gagtggaacg caacctggct gctcctggcc 360 cttggctggr tcttcgtccctgtgtacatc gcagcaggtg tggtcacaat gccgcagtat 420 ctgaagaagc gatttgggggccagaggatc cagrtgtaca tgtctgtcct gtctctcatc 480 ctctacatct tcaccaagatctcgactgac atcttctctg gagccctctt catccagatg 540 gcattgggct ggaacctgtacctctccaca gggatcctgc tggtggtgac tgccgtctac 600 accattgcag gtggcctcatggccgtgatc tacacagatg ctctgcagac ggtgatcatg 660 gtagggggag ccctggtcctcatgtttctg ggctttcagg acgtgggctg gtacccaggc 720 ctggagcagc ggtacaggcaggccatccct aatgtcacag tccccaacac cacctgtcac 780 ctcccacggc ccgatgctttccacatgctt cgggaccctg tgagcgggga catcccttgg 840 ccaggtctca ttttcgggctcacagtgctg gccacctggt gttggtgcac agaccaggtc 900 attgtgcagc ggtctctctcggccaagagt ctgtctcatg ccaagggagg ctccgtgctg 960 gggggctacc tgaagatcctccccatgttc ttcatcgtca tgcctggcat gatcagccgg 1020 gccctgttcc cagacgaggtgggctgcgtg gaccctgatg tctgccaaag aatctgtggg 1080 gcccgagtgg gatgttccaacattgcctac cctaagttgg tcatggccct catgcctgtt 1140 ggtctgcggg ggctgatgattgccgtgatc atggccgctc tcatgagctc actcacctcc 1200 atcttcaaca gcagcagcaccctgttcacc attgatgtgt ggcagcgctt ccgcaggaag 1260 tcaacagagc aggagctgatggtggtgggc agagtgtttg tggtgttcct ggttgtcatc 1320 agcatcctct ggatccccatcatccaaagc tccaacagtg ggcagctctt cgactacatc 1380 caggctgtca ccagttacctggccccaccc atcaccgctc tcttcctgct ggccatcttc 1440 tgcaagaggg tcacagagcccggagctttc tggggcctcg tgtttggcct gggagtgggg 1500 cttctgcgta tgatcctggagttctcatac ccagcgccag cctgtgggga ggtggaccgg 1560 aggccagcag tgctgaaggacttccactac ctgtactttg caatcctcct ctgcgggctc 1620 actgccatcg tcattgtcattgtcagcctc tgtacaactc ccatccctga ggaacagctc 1680 acacgcctca catggtggactcggaactgc cccctctctg agctggagaa ggaggcccac 1740 gagagcacac cggagatatccgagaggcca gccggggagt gccctgcagg aggtggagcg 1800 gcagagaact cgagcctgggccaggagcag cctgaagccc caagcaggtc ctggggaaag 1860 ttgctctgga gctggttctgtgggctctct ggaacaccgg agcaggccct gagcccagca 1920 gagaaggctg cgctagaacagaagctgaca agcattgagg aggagccact ctggagacat 1980 gtctgcaaca tcaatgctgtccttttgctg gccatcaaca tcttcctctg gggctatttt 2040 gcgtga 2046 7 681 PRThomo sapiens VARIANT 124, 152 Xaa = Any Amino Acid 7 Met Ser Lys Glu LeuAla Ala Met Gly Pro Gly Ala Ser Gly Asp Gly 1 5 10 15 Val Arg Thr GluThr Ala Pro His Ile Ala Leu Asp Ser Arg Val Gly 20 25 30 Leu His Ala TyrAsp Ile Ser Val Val Val Ile Tyr Phe Val Phe Val 35 40 45 Ile Ala Val GlyIle Trp Ser Ser Ile Arg Ala Ser Arg Gly Thr Ile 50 55 60 Gly Gly Tyr PheLeu Ala Gly Arg Ser Met Ser Trp Trp Pro Ile Gly 65 70 75 80 Ala Ser LeuMet Ser Ser Asn Val Gly Ser Gly Leu Phe Ile Gly Leu 85 90 95 Ala Gly ThrGly Ala Ala Gly Gly Leu Ala Val Gly Gly Phe Glu Trp 100 105 110 Asn AlaThr Trp Leu Leu Leu Ala Leu Gly Trp Xaa Phe Val Pro Val 115 120 125 TyrIle Ala Ala Gly Val Val Thr Met Pro Gln Tyr Leu Lys Lys Arg 130 135 140Phe Gly Gly Gln Arg Ile Gln Xaa Tyr Met Ser Val Leu Ser Leu Ile 145 150155 160 Leu Tyr Ile Phe Thr Lys Ile Ser Thr Asp Ile Phe Ser Gly Ala Leu165 170 175 Phe Ile Gln Met Ala Leu Gly Trp Asn Leu Tyr Leu Ser Thr GlyIle 180 185 190 Leu Leu Val Val Thr Ala Val Tyr Thr Ile Ala Gly Gly LeuMet Ala 195 200 205 Val Ile Tyr Thr Asp Ala Leu Gln Thr Val Ile Met ValGly Gly Ala 210 215 220 Leu Val Leu Met Phe Leu Gly Phe Gln Asp Val GlyTrp Tyr Pro Gly 225 230 235 240 Leu Glu Gln Arg Tyr Arg Gln Ala Ile ProAsn Val Thr Val Pro Asn 245 250 255 Thr Thr Cys His Leu Pro Arg Pro AspAla Phe His Met Leu Arg Asp 260 265 270 Pro Val Ser Gly Asp Ile Pro TrpPro Gly Leu Ile Phe Gly Leu Thr 275 280 285 Val Leu Ala Thr Trp Cys TrpCys Thr Asp Gln Val Ile Val Gln Arg 290 295 300 Ser Leu Ser Ala Lys SerLeu Ser His Ala Lys Gly Gly Ser Val Leu 305 310 315 320 Gly Gly Tyr LeuLys Ile Leu Pro Met Phe Phe Ile Val Met Pro Gly 325 330 335 Met Ile SerArg Ala Leu Phe Pro Asp Glu Val Gly Cys Val Asp Pro 340 345 350 Asp ValCys Gln Arg Ile Cys Gly Ala Arg Val Gly Cys Ser Asn Ile 355 360 365 AlaTyr Pro Lys Leu Val Met Ala Leu Met Pro Val Gly Leu Arg Gly 370 375 380Leu Met Ile Ala Val Ile Met Ala Ala Leu Met Ser Ser Leu Thr Ser 385 390395 400 Ile Phe Asn Ser Ser Ser Thr Leu Phe Thr Ile Asp Val Trp Gln Arg405 410 415 Phe Arg Arg Lys Ser Thr Glu Gln Glu Leu Met Val Val Gly ArgVal 420 425 430 Phe Val Val Phe Leu Val Val Ile Ser Ile Leu Trp Ile ProIle Ile 435 440 445 Gln Ser Ser Asn Ser Gly Gln Leu Phe Asp Tyr Ile GlnAla Val Thr 450 455 460 Ser Tyr Leu Ala Pro Pro Ile Thr Ala Leu Phe LeuLeu Ala Ile Phe 465 470 475 480 Cys Lys Arg Val Thr Glu Pro Gly Ala PheTrp Gly Leu Val Phe Gly 485 490 495 Leu Gly Val Gly Leu Leu Arg Met IleLeu Glu Phe Ser Tyr Pro Ala 500 505 510 Pro Ala Cys Gly Glu Val Asp ArgArg Pro Ala Val Leu Lys Asp Phe 515 520 525 His Tyr Leu Tyr Phe Ala IleLeu Leu Cys Gly Leu Thr Ala Ile Val 530 535 540 Ile Val Ile Val Ser LeuCys Thr Thr Pro Ile Pro Glu Glu Gln Leu 545 550 555 560 Thr Arg Leu ThrTrp Trp Thr Arg Asn Cys Pro Leu Ser Glu Leu Glu 565 570 575 Lys Glu AlaHis Glu Ser Thr Pro Glu Ile Ser Glu Arg Pro Ala Gly 580 585 590 Glu CysPro Ala Gly Gly Gly Ala Ala Glu Asn Ser Ser Leu Gly Gln 595 600 605 GluGln Pro Glu Ala Pro Ser Arg Ser Trp Gly Lys Leu Leu Trp Ser 610 615 620Trp Phe Cys Gly Leu Ser Gly Thr Pro Glu Gln Ala Leu Ser Pro Ala 625 630635 640 Glu Lys Ala Ala Leu Glu Gln Lys Leu Thr Ser Ile Glu Glu Glu Pro645 650 655 Leu Trp Arg His Val Cys Asn Ile Asn Ala Val Leu Leu Leu AlaIle 660 665 670 Asn Ile Phe Leu Trp Gly Tyr Phe Ala 675 680 8 2025 DNAhomo sapiens 8 atggggcctg gagcttcagg ggacggggtc aggactgaga cagctccacacatagcactg 60 gactccagag ttggtctgca cgcctacgac atcagcgtgg tggtcatctactttgtcttc 120 gtcattgctg tggggatctg gtcgtccatc cgtgcaagtc gagggaccattggcggctat 180 ttcctggccg ggaggtccat gagctggtgg ccaattggag catctctgatgtccagcaat 240 gtgggcagtg gcttgttcat cggcctggct gggacagggg ctgccggaggccttgccgta 300 ggtggcttcg agtggaacgc aacctggctg ctcctggccc ttggctgggtcttcgtccct 360 gtgtacatcg cagcaggtgt ggtcacaatg ccgcagtatc tgaagaagcgatttgggggc 420 cagaggatcc aggtgtacat gtctgtcctg tctctcatcc tctacatcttcaccaagatc 480 tcgactgaca tcttctctgg agccctcttc atccagatgg cattgggctggaacctgtac 540 ctctccacag ggatcctgct ggtggtgact gccgtctaca ccattgcaggtggcctcatg 600 gccgtgatct acacagatgc tctgcagacg gtgatcatgg tagggggagccctggtcctc 660 atgtttctgg gctttcagga cgtgggctgg tacccaggcc tggagcagcggtacaggcag 720 gccatcccta atgtcacagt ccccaacacc acctgtcacc tcccacggcccgatgctttc 780 cacatgcttc gggaccctgt gagcggggac atcccttggc caggtctcattttcgggctc 840 acagtgctgg ccacctggtg ttggtgcaca gaccaggtca ttgtgcagcggtctctctcg 900 gccaagagtc tgtctcatgc caagggaggc tccgtgctgg ggggctacctgaagatcctc 960 cccatgttct tcatcgtcat gcctggcatg atcagccggg ccctgttcccagacgaggtg 1020 ggctgcgtgg accctgatgt ctgccaaaga atctgtgggg cccgagtgggatgttccaac 1080 attgcctacc ctaagttggt catggccctc atgcctgttg gtctgcgggggctgatgatt 1140 gccgtgatca tggccgctct catgagctca ctcacctcca tcttcaacagcagcagcacc 1200 ctgttcacca ttgatgtgtg gcagcgcttc cgcaggaagt caacagagcaggagctgatg 1260 gtggtgggca gagtgtttgt ggtgttcctg gttgtcatca gcatcctctggatccccatc 1320 atccaaagct ccaacagtgg gcagctcttc gactacatcc aggctgtcaccagttacctg 1380 gccccaccca tcaccgctct cttcctgctg gccatcttct gcaagagggtcacagagccc 1440 ggagctttct ggggcctcgt gtttggcctg ggagtggggc ttctgcgtatgatcctggag 1500 ttctcatacc cagcgccagc ctgtggggag gtggaccgga ggccagcagtgctgaaggac 1560 ttccactacc tgtactttgc aatcctcctc tgcgggctca ctgccatcgtcattgtcatt 1620 gtcagcctct gtacaactcc catccctgag gaacagctca cacgcctcacatggtggact 1680 cggaactgcc ccctctctga gctggagaag gaggcccacg agagcacaccggagatatcc 1740 gagaggccag ccggggagtg ccctgcagga ggtggagcgg cagagaactcgagcctgggc 1800 caggagcagc ctgaagcccc aagcaggtcc tggggaaagt tgctctggagctggttctgt 1860 gggctctctg gaacaccgga gcaggccctg agcccagcag agaaggctgcgctagaacag 1920 aagctgacaa gcattgagga ggagccactc tggagacatg tctgcaacatcaatgctgtc 1980 cttttgctgg ccatcaacat cttcctctgg ggctattttg cgtga 2025 9674 PRT homo sapiens 9 Met Gly Pro Gly Ala Ser Gly Asp Gly Val Arg ThrGlu Thr Ala Pro 1 5 10 15 His Ile Ala Leu Asp Ser Arg Val Gly Leu HisAla Tyr Asp Ile Ser 20 25 30 Val Val Val Ile Tyr Phe Val Phe Val Ile AlaVal Gly Ile Trp Ser 35 40 45 Ser Ile Arg Ala Ser Arg Gly Thr Ile Gly GlyTyr Phe Leu Ala Gly 50 55 60 Arg Ser Met Ser Trp Trp Pro Ile Gly Ala SerLeu Met Ser Ser Asn 65 70 75 80 Val Gly Ser Gly Leu Phe Ile Gly Leu AlaGly Thr Gly Ala Ala Gly 85 90 95 Gly Leu Ala Val Gly Gly Phe Glu Trp AsnAla Thr Trp Leu Leu Leu 100 105 110 Ala Leu Gly Trp Val Phe Val Pro ValTyr Ile Ala Ala Gly Val Val 115 120 125 Thr Met Pro Gln Tyr Leu Lys LysArg Phe Gly Gly Gln Arg Ile Gln 130 135 140 Val Tyr Met Ser Val Leu SerLeu Ile Leu Tyr Ile Phe Thr Lys Ile 145 150 155 160 Ser Thr Asp Ile PheSer Gly Ala Leu Phe Ile Gln Met Ala Leu Gly 165 170 175 Trp Asn Leu TyrLeu Ser Thr Gly Ile Leu Leu Val Val Thr Ala Val 180 185 190 Tyr Thr IleAla Gly Gly Leu Met Ala Val Ile Tyr Thr Asp Ala Leu 195 200 205 Gln ThrVal Ile Met Val Gly Gly Ala Leu Val Leu Met Phe Leu Gly 210 215 220 PheGln Asp Val Gly Trp Tyr Pro Gly Leu Glu Gln Arg Tyr Arg Gln 225 230 235240 Ala Ile Pro Asn Val Thr Val Pro Asn Thr Thr Cys His Leu Pro Arg 245250 255 Pro Asp Ala Phe His Met Leu Arg Asp Pro Val Ser Gly Asp Ile Pro260 265 270 Trp Pro Gly Leu Ile Phe Gly Leu Thr Val Leu Ala Thr Trp CysTrp 275 280 285 Cys Thr Asp Gln Val Ile Val Gln Arg Ser Leu Ser Ala LysSer Leu 290 295 300 Ser His Ala Lys Gly Gly Ser Val Leu Gly Gly Tyr LeuLys Ile Leu 305 310 315 320 Pro Met Phe Phe Ile Val Met Pro Gly Met IleSer Arg Ala Leu Phe 325 330 335 Pro Asp Glu Val Gly Cys Val Asp Pro AspVal Cys Gln Arg Ile Cys 340 345 350 Gly Ala Arg Val Gly Cys Ser Asn IleAla Tyr Pro Lys Leu Val Met 355 360 365 Ala Leu Met Pro Val Gly Leu ArgGly Leu Met Ile Ala Val Ile Met 370 375 380 Ala Ala Leu Met Ser Ser LeuThr Ser Ile Phe Asn Ser Ser Ser Thr 385 390 395 400 Leu Phe Thr Ile AspVal Trp Gln Arg Phe Arg Arg Lys Ser Thr Glu 405 410 415 Gln Glu Leu MetVal Val Gly Arg Val Phe Val Val Phe Leu Val Val 420 425 430 Ile Ser IleLeu Trp Ile Pro Ile Ile Gln Ser Ser Asn Ser Gly Gln 435 440 445 Leu PheAsp Tyr Ile Gln Ala Val Thr Ser Tyr Leu Ala Pro Pro Ile 450 455 460 ThrAla Leu Phe Leu Leu Ala Ile Phe Cys Lys Arg Val Thr Glu Pro 465 470 475480 Gly Ala Phe Trp Gly Leu Val Phe Gly Leu Gly Val Gly Leu Leu Arg 485490 495 Met Ile Leu Glu Phe Ser Tyr Pro Ala Pro Ala Cys Gly Glu Val Asp500 505 510 Arg Arg Pro Ala Val Leu Lys Asp Phe His Tyr Leu Tyr Phe AlaIle 515 520 525 Leu Leu Cys Gly Leu Thr Ala Ile Val Ile Val Ile Val SerLeu Cys 530 535 540 Thr Thr Pro Ile Pro Glu Glu Gln Leu Thr Arg Leu ThrTrp Trp Thr 545 550 555 560 Arg Asn Cys Pro Leu Ser Glu Leu Glu Lys GluAla His Glu Ser Thr 565 570 575 Pro Glu Ile Ser Glu Arg Pro Ala Gly GluCys Pro Ala Gly Gly Gly 580 585 590 Ala Ala Glu Asn Ser Ser Leu Gly GlnGlu Gln Pro Glu Ala Pro Ser 595 600 605 Arg Ser Trp Gly Lys Leu Leu TrpSer Trp Phe Cys Gly Leu Ser Gly 610 615 620 Thr Pro Glu Gln Ala Leu SerPro Ala Glu Lys Ala Ala Leu Glu Gln 625 630 635 640 Lys Leu Thr Ser IleGlu Glu Glu Pro Leu Trp Arg His Val Cys Asn 645 650 655 Ile Asn Ala ValLeu Leu Leu Ala Ile Asn Ile Phe Leu Trp Gly Tyr 660 665 670 Phe Ala 102238 DNA homo sapiens 10 atgagcaagg agctggcagc aatggggcct ggagcttcaggggacggggt caggactgag 60 acagctccac acatagcact ggactccaga gttggtctgcacgcctacga catcagcgtg 120 gtggtcatct actttgtctt cgtcattgct gtggggatctggtcgtccat ccgtgcaagt 180 cgagggacca ttggcggcta tttcctggcc gggaggtccatgagctggtg gccaattgga 240 gcatctctga tgtccagcaa tgtgggcagt ggcttgttcatcggcctggc tgggacaggg 300 gctgccggag gccttgccgt aggtggcttc gagtggaacatgaggaaatc aaggtctgga 360 ggagacagag ggatccatcc aaggtcacac gggaggactggggtcaggtc ccaggtctct 420 tatttctctg ttcgggggcc tcccacagca cagcactgcctctgggtggg aagccgcccc 480 tctgtctaca tccaggacct ggataccttc ttcttctccccactctccca ggcaacctgg 540 ctgctcctgg cccttggctg ggtcttcgtc cctgtgtacatcgcagcagg tgtggtcaca 600 atgccgcagt atctgaagaa gcgatttggg ggccagaggatccaggtgta catgtctgtc 660 ctgtctctca tcctctacat cttcaccaag atctcgactgacatcttctc tggagccctc 720 ttcatccaga tggcattggg ctggaacctg tacctctccacagggatcct gctggtggtg 780 actgccgtct acaccattgc aggtggcctc atggccgtgatctacacaga tgctctgcag 840 acggtgatca tggtaggggg agccctggtc ctcatgtttctgggctttca ggacgtgggc 900 tggtacccag gcctggagca gcggtacagg caggccatccctaatgtcac agtccccaac 960 accacctgtc acctcccacg gcccgatgct ttccacatgcttcgggaccc tgtgagyggg 1020 gacatccctt ggccaggtct cattttcggg ctcacagtgctggccacctg gtgttggtgc 1080 acagaccagg tcattgtgca gcggtctctc tcggccaagagtctgtctca tgccaaggga 1140 ggctccgtgc tggggggcta cctgaagatc ctccccatgttcttcatcgt catgcctggc 1200 atgatcagcc gggccctgtt cccagacgag gtgggctgcgtggaccctga tgtctgccaa 1260 agaatctgtg gggcccgagt gggatgttcc aacattgcctaccctaagtt ggtcatggcc 1320 ctcatgcctg ttggtctgcg ggggctgatg attgccgtgatcatggccgc tctcatgagc 1380 tcactcacct ccatcttcaa cagcagcagc accctgttcaccattgatgt gtggcagcgc 1440 ttccgcagga agtcaacaga gcaggagctg atggtggtgggcagagtgtt tgtggtgttc 1500 ctggttgtca tcagcatcct ctggatcccc atcatccaaagctccaacag tgggcagctc 1560 ttcgactaca tccaggctgt caccagttac ctggccccacccatcaccgc tctcttcctg 1620 ctggccatct tctgcaagag ggtcacagag cccggagctttctggggcct cgtgtttggc 1680 ctgggagtgg ggcttctgcg tatgatcctg gagttctcatacccagcgcc agcctgtggg 1740 gaggtggacc ggaggccagc agtgctgaag gacttccactacctgtactt tgcaatcctc 1800 ctctgcgggc tcactgccat cgtcattgtc attgtcagcctctgtacaac tcccatccct 1860 gaggaacagc tcacacgcct cacatggtgg actcggaactgccccctctc tgagctggag 1920 aaggaggccc acgagagcac accggagata tccgagaggccagccgggga gtgccctgca 1980 ggaggtggag cggcagagaa ctcgagcctg ggccaggagcagcctgaagc cccaagcagg 2040 tcctggggaa agttgctctg gagctggttc tgtgggctctctggaacacc ggagcaggcc 2100 ctgagcccag cagagaaggc tgcgctagaa cagaagctgacaagcattga ggaggagcca 2160 ctctggagac atgtctgcaa catcaatgct gtccttttgctggccatcaa catcttcctc 2220 tggggctatt ttgcgtga 2238 11 745 PRT homosapiens 11 Met Ser Lys Glu Leu Ala Ala Met Gly Pro Gly Ala Ser Gly AspGly 1 5 10 15 Val Arg Thr Glu Thr Ala Pro His Ile Ala Leu Asp Ser ArgVal Gly 20 25 30 Leu His Ala Tyr Asp Ile Ser Val Val Val Ile Tyr Phe ValPhe Val 35 40 45 Ile Ala Val Gly Ile Trp Ser Ser Ile Arg Ala Ser Arg GlyThr Ile 50 55 60 Gly Gly Tyr Phe Leu Ala Gly Arg Ser Met Ser Trp Trp ProIle Gly 65 70 75 80 Ala Ser Leu Met Ser Ser Asn Val Gly Ser Gly Leu PheIle Gly Leu 85 90 95 Ala Gly Thr Gly Ala Ala Gly Gly Leu Ala Val Gly GlyPhe Glu Trp 100 105 110 Asn Met Arg Lys Ser Arg Ser Gly Gly Asp Arg GlyIle His Pro Arg 115 120 125 Ser His Gly Arg Thr Gly Val Arg Ser Gln ValSer Tyr Phe Ser Val 130 135 140 Arg Gly Pro Pro Thr Ala Gln His Cys LeuTrp Val Gly Ser Arg Pro 145 150 155 160 Ser Val Tyr Ile Gln Asp Leu AspThr Phe Phe Phe Ser Pro Leu Ser 165 170 175 Gln Ala Thr Trp Leu Leu LeuAla Leu Gly Trp Val Phe Val Pro Val 180 185 190 Tyr Ile Ala Ala Gly ValVal Thr Met Pro Gln Tyr Leu Lys Lys Arg 195 200 205 Phe Gly Gly Gln ArgIle Gln Val Tyr Met Ser Val Leu Ser Leu Ile 210 215 220 Leu Tyr Ile PheThr Lys Ile Ser Thr Asp Ile Phe Ser Gly Ala Leu 225 230 235 240 Phe IleGln Met Ala Leu Gly Trp Asn Leu Tyr Leu Ser Thr Gly Ile 245 250 255 LeuLeu Val Val Thr Ala Val Tyr Thr Ile Ala Gly Gly Leu Met Ala 260 265 270Val Ile Tyr Thr Asp Ala Leu Gln Thr Val Ile Met Val Gly Gly Ala 275 280285 Leu Val Leu Met Phe Leu Gly Phe Gln Asp Val Gly Trp Tyr Pro Gly 290295 300 Leu Glu Gln Arg Tyr Arg Gln Ala Ile Pro Asn Val Thr Val Pro Asn305 310 315 320 Thr Thr Cys His Leu Pro Arg Pro Asp Ala Phe His Met LeuArg Asp 325 330 335 Pro Val Ser Gly Asp Ile Pro Trp Pro Gly Leu Ile PheGly Leu Thr 340 345 350 Val Leu Ala Thr Trp Cys Trp Cys Thr Asp Gln ValIle Val Gln Arg 355 360 365 Ser Leu Ser Ala Lys Ser Leu Ser His Ala LysGly Gly Ser Val Leu 370 375 380 Gly Gly Tyr Leu Lys Ile Leu Pro Met PhePhe Ile Val Met Pro Gly 385 390 395 400 Met Ile Ser Arg Ala Leu Phe ProAsp Glu Val Gly Cys Val Asp Pro 405 410 415 Asp Val Cys Gln Arg Ile CysGly Ala Arg Val Gly Cys Ser Asn Ile 420 425 430 Ala Tyr Pro Lys Leu ValMet Ala Leu Met Pro Val Gly Leu Arg Gly 435 440 445 Leu Met Ile Ala ValIle Met Ala Ala Leu Met Ser Ser Leu Thr Ser 450 455 460 Ile Phe Asn SerSer Ser Thr Leu Phe Thr Ile Asp Val Trp Gln Arg 465 470 475 480 Phe ArgArg Lys Ser Thr Glu Gln Glu Leu Met Val Val Gly Arg Val 485 490 495 PheVal Val Phe Leu Val Val Ile Ser Ile Leu Trp Ile Pro Ile Ile 500 505 510Gln Ser Ser Asn Ser Gly Gln Leu Phe Asp Tyr Ile Gln Ala Val Thr 515 520525 Ser Tyr Leu Ala Pro Pro Ile Thr Ala Leu Phe Leu Leu Ala Ile Phe 530535 540 Cys Lys Arg Val Thr Glu Pro Gly Ala Phe Trp Gly Leu Val Phe Gly545 550 555 560 Leu Gly Val Gly Leu Leu Arg Met Ile Leu Glu Phe Ser TyrPro Ala 565 570 575 Pro Ala Cys Gly Glu Val Asp Arg Arg Pro Ala Val LeuLys Asp Phe 580 585 590 His Tyr Leu Tyr Phe Ala Ile Leu Leu Cys Gly LeuThr Ala Ile Val 595 600 605 Ile Val Ile Val Ser Leu Cys Thr Thr Pro IlePro Glu Glu Gln Leu 610 615 620 Thr Arg Leu Thr Trp Trp Thr Arg Asn CysPro Leu Ser Glu Leu Glu 625 630 635 640 Lys Glu Ala His Glu Ser Thr ProGlu Ile Ser Glu Arg Pro Ala Gly 645 650 655 Glu Cys Pro Ala Gly Gly GlyAla Ala Glu Asn Ser Ser Leu Gly Gln 660 665 670 Glu Gln Pro Glu Ala ProSer Arg Ser Trp Gly Lys Leu Leu Trp Ser 675 680 685 Trp Phe Cys Gly LeuSer Gly Thr Pro Glu Gln Ala Leu Ser Pro Ala 690 695 700 Glu Lys Ala AlaLeu Glu Gln Lys Leu Thr Ser Ile Glu Glu Glu Pro 705 710 715 720 Leu TrpArg His Val Cys Asn Ile Asn Ala Val Leu Leu Leu Ala Ile 725 730 735 AsnIle Phe Leu Trp Gly Tyr Phe Ala 740 745 12 2217 DNA homo sapiens 12atggggcctg gagcttcagg ggacggggtc aggactgaga cagctccaca catagcactg 60gactccagag ttggtctgca cgcctacgac atcagcgtgg tggtcatcta ctttgtcttc 120gtcattgctg tggggatctg gtcgtccatc cgtgcaagtc gagggaccat tggcggctat 180ttcctggccg ggaggtccat gagctggtgg ccaattggag catctctgat gtccagcaat 240gtgggcagtg gcttgttcat cggcctggct gggacagggg ctgccggagg ccttgccgta 300ggtggcttcg agtggaacat gaggaaatca aggtctggag gagacagagg gatccatcca 360aggtcacacg ggaggactgg ggtcaggtcc caggtctctt atttctctgt tcgggggcct 420cccacagcac agcactgcct ctgggtggga agccgcccct ctgtctacat ccaggacctg 480gataccttct tcttctcccc actctcccag gcaacctggc tgctcctggc ccttggctgg 540gtcttcgtcc ctgtgtacat cgcagcaggt gtggtcacaa tgccgcagta tctgaagaag 600cgatttgggg gccagaggat ccaggtgtac atgtctgtcc tgtctctcat cctctacatc 660ttcaccaaga tctcgactga catcttctct ggagccctct tcatccagat ggcattgggc 720tggaacctgt acctctccac agggatcctg ctggtggtga ctgccgtcta caccattgca 780ggtggcctca tggccgtgat ctacacagat gctctgcaga cggtgatcat ggtaggggga 840gccctggtcc tcatgtttct gggctttcag gacgtgggct ggtacccagg cctggagcag 900cggtacaggc aggccatccc taatgtcaca gtccccaaca ccacctgtca cctcccacgg 960cccgatgctt tccacatgct tcgggaccct gtgagygggg acatcccttg gccaggtctc 1020attttcgggc tcacagtgct ggccacctgg tgttggtgca cagaccaggt cattgtgcag 1080cggtctctct cggccaagag tctgtctcat gccaagggag gctccgtgct ggggggctac 1140ctgaagatcc tccccatgtt cttcatcgtc atgcctggca tgatcagccg ggccctgttc 1200ccagacgagg tgggctgcgt ggaccctgat gtctgccaaa gaatctgtgg ggcccgagtg 1260ggatgttcca acattgccta ccctaagttg gtcatggccc tcatgcctgt tggtctgcgg 1320gggctgatga ttgccgtgat catggccgct ctcatgagct cactcacctc catcttcaac 1380agcagcagca ccctgttcac cattgatgtg tggcagcgct tccgcaggaa gtcaacagag 1440caggagctga tggtggtggg cagagtgttt gtggtgttcc tggttgtcat cagcatcctc 1500tggatcccca tcatccaaag ctccaacagt gggcagctct tcgactacat ccaggctgtc 1560accagttacc tggccccacc catcaccgct ctcttcctgc tggccatctt ctgcaagagg 1620gtcacagagc ccggagcttt ctggggcctc gtgtttggcc tgggagtggg gcttctgcgt 1680atgatcctgg agttctcata cccagcgcca gcctgtgggg aggtggaccg gaggccagca 1740gtgctgaagg acttccacta cctgtacttt gcaatcctcc tctgcgggct cactgccatc 1800gtcattgtca ttgtcagcct ctgtacaact cccatccctg aggaacagct cacacgcctc 1860acatggtgga ctcggaactg ccccctctct gagctggaga aggaggccca cgagagcaca 1920ccggagatat ccgagaggcc agccggggag tgccctgcag gaggtggagc ggcagagaac 1980tcgagcctgg gccaggagca gcctgaagcc ccaagcaggt cctggggaaa gttgctctgg 2040agctggttct gtgggctctc tggaacaccg gagcaggccc tgagcccagc agagaaggct 2100gcgctagaac agaagctgac aagcattgag gaggagccac tctggagaca tgtctgcaac 2160atcaatgctg tccttttgct ggccatcaac atcttcctct ggggctattt tgcgtga 2217 13738 PRT homo sapiens 13 Met Gly Pro Gly Ala Ser Gly Asp Gly Val Arg ThrGlu Thr Ala Pro 1 5 10 15 His Ile Ala Leu Asp Ser Arg Val Gly Leu HisAla Tyr Asp Ile Ser 20 25 30 Val Val Val Ile Tyr Phe Val Phe Val Ile AlaVal Gly Ile Trp Ser 35 40 45 Ser Ile Arg Ala Ser Arg Gly Thr Ile Gly GlyTyr Phe Leu Ala Gly 50 55 60 Arg Ser Met Ser Trp Trp Pro Ile Gly Ala SerLeu Met Ser Ser Asn 65 70 75 80 Val Gly Ser Gly Leu Phe Ile Gly Leu AlaGly Thr Gly Ala Ala Gly 85 90 95 Gly Leu Ala Val Gly Gly Phe Glu Trp AsnMet Arg Lys Ser Arg Ser 100 105 110 Gly Gly Asp Arg Gly Ile His Pro ArgSer His Gly Arg Thr Gly Val 115 120 125 Arg Ser Gln Val Ser Tyr Phe SerVal Arg Gly Pro Pro Thr Ala Gln 130 135 140 His Cys Leu Trp Val Gly SerArg Pro Ser Val Tyr Ile Gln Asp Leu 145 150 155 160 Asp Thr Phe Phe PheSer Pro Leu Ser Gln Ala Thr Trp Leu Leu Leu 165 170 175 Ala Leu Gly TrpVal Phe Val Pro Val Tyr Ile Ala Ala Gly Val Val 180 185 190 Thr Met ProGln Tyr Leu Lys Lys Arg Phe Gly Gly Gln Arg Ile Gln 195 200 205 Val TyrMet Ser Val Leu Ser Leu Ile Leu Tyr Ile Phe Thr Lys Ile 210 215 220 SerThr Asp Ile Phe Ser Gly Ala Leu Phe Ile Gln Met Ala Leu Gly 225 230 235240 Trp Asn Leu Tyr Leu Ser Thr Gly Ile Leu Leu Val Val Thr Ala Val 245250 255 Tyr Thr Ile Ala Gly Gly Leu Met Ala Val Ile Tyr Thr Asp Ala Leu260 265 270 Gln Thr Val Ile Met Val Gly Gly Ala Leu Val Leu Met Phe LeuGly 275 280 285 Phe Gln Asp Val Gly Trp Tyr Pro Gly Leu Glu Gln Arg TyrArg Gln 290 295 300 Ala Ile Pro Asn Val Thr Val Pro Asn Thr Thr Cys HisLeu Pro Arg 305 310 315 320 Pro Asp Ala Phe His Met Leu Arg Asp Pro ValSer Gly Asp Ile Pro 325 330 335 Trp Pro Gly Leu Ile Phe Gly Leu Thr ValLeu Ala Thr Trp Cys Trp 340 345 350 Cys Thr Asp Gln Val Ile Val Gln ArgSer Leu Ser Ala Lys Ser Leu 355 360 365 Ser His Ala Lys Gly Gly Ser ValLeu Gly Gly Tyr Leu Lys Ile Leu 370 375 380 Pro Met Phe Phe Ile Val MetPro Gly Met Ile Ser Arg Ala Leu Phe 385 390 395 400 Pro Asp Glu Val GlyCys Val Asp Pro Asp Val Cys Gln Arg Ile Cys 405 410 415 Gly Ala Arg ValGly Cys Ser Asn Ile Ala Tyr Pro Lys Leu Val Met 420 425 430 Ala Leu MetPro Val Gly Leu Arg Gly Leu Met Ile Ala Val Ile Met 435 440 445 Ala AlaLeu Met Ser Ser Leu Thr Ser Ile Phe Asn Ser Ser Ser Thr 450 455 460 LeuPhe Thr Ile Asp Val Trp Gln Arg Phe Arg Arg Lys Ser Thr Glu 465 470 475480 Gln Glu Leu Met Val Val Gly Arg Val Phe Val Val Phe Leu Val Val 485490 495 Ile Ser Ile Leu Trp Ile Pro Ile Ile Gln Ser Ser Asn Ser Gly Gln500 505 510 Leu Phe Asp Tyr Ile Gln Ala Val Thr Ser Tyr Leu Ala Pro ProIle 515 520 525 Thr Ala Leu Phe Leu Leu Ala Ile Phe Cys Lys Arg Val ThrGlu Pro 530 535 540 Gly Ala Phe Trp Gly Leu Val Phe Gly Leu Gly Val GlyLeu Leu Arg 545 550 555 560 Met Ile Leu Glu Phe Ser Tyr Pro Ala Pro AlaCys Gly Glu Val Asp 565 570 575 Arg Arg Pro Ala Val Leu Lys Asp Phe HisTyr Leu Tyr Phe Ala Ile 580 585 590 Leu Leu Cys Gly Leu Thr Ala Ile ValIle Val Ile Val Ser Leu Cys 595 600 605 Thr Thr Pro Ile Pro Glu Glu GlnLeu Thr Arg Leu Thr Trp Trp Thr 610 615 620 Arg Asn Cys Pro Leu Ser GluLeu Glu Lys Glu Ala His Glu Ser Thr 625 630 635 640 Pro Glu Ile Ser GluArg Pro Ala Gly Glu Cys Pro Ala Gly Gly Gly 645 650 655 Ala Ala Glu AsnSer Ser Leu Gly Gln Glu Gln Pro Glu Ala Pro Ser 660 665 670 Arg Ser TrpGly Lys Leu Leu Trp Ser Trp Phe Cys Gly Leu Ser Gly 675 680 685 Thr ProGlu Gln Ala Leu Ser Pro Ala Glu Lys Ala Ala Leu Glu Gln 690 695 700 LysLeu Thr Ser Ile Glu Glu Glu Pro Leu Trp Arg His Val Cys Asn 705 710 715720 Ile Asn Ala Val Leu Leu Leu Ala Ile Asn Ile Phe Leu Trp Gly Tyr 725730 735 Phe Ala

What is claimed is:
 1. An isolated nucleic acid molecule comprising thenucleotide sequence of SEQ ID NO: 1 or SEQ ID NO:
 2. 2. An isolatednucleic acid molecule comprising a nucleotide sequence that: (a) encodesthe amino acid sequence shown in SEQ ID NO: 3 or SEQ ID NO: 4; and (b)hybridizes under highly stringent conditions to the nucleotide sequenceof SEQ ID NO: 1 or SEQ ID NO: 2, or the complement thereof.
 3. Anisolated nucleic acid molecule comprising a nucleotide sequence thatencodes the amino acid sequence shown in SEQ ID NO: 3 or SEQ ID NO: 4.4. A recombinant expression vector comprising the isolated nucleic acidmolecule of claim
 1. 5. A host cell comprising the recombinantexpression vector of claim
 4. 6. A substantially isolated protein havingthe activity of the protein shown in SEQ ID NOS: 3 or 4, which isencoded by a nucleotide sequence that hybridizes to the complement ofSEQ ID NO: 1 or SEQ ID NO: 2 under highly stringent conditions.
 7. Anisolated nucleic acid molecule comprising the nucleotide sequence of SEQID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10 or SEQ ID NO:
 12. 8. An isolatednucleic acid molecule comprising a nucleotide sequence that: (c) encodesthe amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 orSEQ ID NO: 13; and (d) hybridizes under highly stringent conditions tothe nucleotide sequence of SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10 orSEQ ID NO: 12, or the complement thereof.
 9. An isolated nucleic acidmolecule comprising a nucleotide sequence that encodes the amino acidsequence shown in SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ IDNO:
 13. 10. A recombinant expression vector comprising the isolatednucleic acid molecule of claim
 7. 11. A host cell comprising therecombinant expression vector of claim
 10. 12. The host cell of claim11, wherein said cell is procaryotic.
 13. The host cell of claim 11,wherein said cell is eucaryotic.
 14. A substantially isolated proteinhaving the activity of the protein shown in SEQ ID NO: 7, SEQ ID NO: 9,SEQ ID NO: 11 or SEQ ID NO: 13, which is encoded by a nucleotidesequence that hybridizes to the complement of SEQ ID NO: 6, SEQ ID NO:8, SEQ ID NO: 10 or SEQ ID NO: 12 under highly stringent conditions.